Theobroma Grandiflorum Research Articles

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

Optimize the production of Aspergillus brasiliensis endoglucanase in a solid-phase bioprocess using cupuaçu shell as substrate. 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 7.35Ug-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 40.50Ug-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 48h, it was possible to observe the production of reducing sugars. 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. 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.

Process optimization for simultaneous production of phenolic acids and enzymes with high transfructosylation activity in cupuassu (Theobroma grandiflorum) residue by submerged fermentation with Aspergillus carbonarius.

The online version contains supplementary material available at 10.1007/s13197-022-05418-z.

Biotechnological methods for pulping copoazú (Theobroma grandiflorum (Wild. ex Spring) Schum) seed and their influence on the fermentation stage

Copoazú is a fruit originated from Colombian Amazon belonging to Theobroma genus, which includes species such as cacao raw material for chocolate. Both species presented similar processing stages, with different process parameters due to the physical qualities of each fruit. Enzymatic, mechanical, and combined methods for pulping copoazú seeds and their effect on the biochemical dynamics of fermentation were evaluated. The pulping treatments evaluated were manual (Ma), mechanical (Me), enzymatic (Ez), and enzymatic-mechanical (Ez-Me), considering the yield as a response variable. The results showed higher percentages of pulp recovery for enzymatic, enzymatic-mechanical, and manual treatments with 61.01%, 66.80%, and 65.71%, respectively. The dynamics of the fermentation showed a reduction of seven to four days for the fermentation stage with the enzymatic treatments, registering up to 85.3% of well-fermented grain at the end of this phase.

Cupuassu (Theobroma grandiflorum [Willd. ex Sprengel] Schumann) Fruit Development: Key Genes Involved in Primary Metabolism and Stress Response

Cupuassu (Theobroma grandiflorum [Willd. ex Sprengel] Schumann) seeds constitute the raw material for oil extraction and fabrication of cupulate (product similar to chocolate). However, fungal diseases such as witches’ broom caused by Moniliophthora perniciosa have interfered with the large-scale development of cupuassu plantations. Cupuassu genetic breeding programmes focus on a variety of biotechnological tools or approaches to select genes related to quality or resistance mechanisms. In this study, we used expression and interactomics analyses of preselected genes involved in fruit quality and/or resistance to better understand the molecular and physiological mechanisms associated with these plant processes. It was found that (i) resistant and susceptible cupuassu genotypes showed different pulp characteristics as well as gene expression patterns; (ii) monosaccharide and carbohydrate transport pathways were enhanced during fruit maturation; (iii) sugar accumulation participated in signal transduction associated with fruit development and stress response in maturing fruits; and (iv) maturing pulp and seeds showed increased phospholipid metabolism and translocation, as well as immune system activation. The TgSTP1, TgWRKY33, TgCZF1, and TgUBA1 genes in cupuassu and the orthologues of DIN10, CNI1, and TET8 identified by the interactomics approach may be good candidates for marker-assisted selection in breeding programmes focusing on both fruit quality and resistance/tolerance to biotic/abiotic stress.

Evaluating the bioenergy potential of cupuassu shell through pyrolysis kinetics, thermodynamic parameters of activation, and evolved gas analysis with TG/FTIR technique

This research aimed to investigate the physicochemical properties and the pyrolysis performance of cupuassu shell (Theobroma grandiflorum) under a thermogravimetric scale. The pyrolysis experiments were carried out using a pure nitrogen atmosphere and at five heating rates (5, 10, 20, 30, and 40 K min−1). The pyrolysis kinetic triplet was calculated from the DTG thermograms using the isoconversional methods of Friedman, Flynn–Wall–Ozawa, Kissinger–Akahira–Sunose, and Starink combined with the compensation effect and master plots methods. TGA analyses revealed that cupuassu shell pyrolysis follows a devolatilization process of two steps: (1) represented by the two-dimensional diffusion-reaction model (D2), with average activation energies within 102.9–107.5 kJ mol−1, and (2) represented by the fourth-order reaction model (F4) with average activation energies within 176.7–243.0 kJ mol−1. Still, the pre-exponential factor value for the first kinetic stage (1.64×108 min−1) was lower than that of the second kinetic stage (3.19×1010 min−1). As verified, the kinetic triplets efficiently derived the global kinetic expression of cupuassu shell pyrolysis, reproducing reliable data on conversion rates. The evaluation of thermodynamic parameters of activation showed that cupuassu shell is viable biomass for pyrolysis applications, with values of ΔH≠ > 0, ΔG≠ > 0, and ΔS≠ < 0. The TGA-FTIR analysis showed that the evolved pyrolysis products were dominated by alcohols, aldehydes, ketones, organic acids, and aromatic/aliphatic hydrocarbons, supporting the utility of cupuassu shell as a potential source for bio-based chemicals production. In summary, the insights of this study validate the cupuassu shell as a prospective feedstock for producing bioenergy and bio-based chemicals, also bringing useful information for engineering purposes in the design or simulation of large-scale pyrolysis reactors.

Enhanced solar photo-electro-Fenton by Theobroma grandiflorum addition during pharmaceuticals elimination in municipal wastewater: Action routes, process improvement, and biodegradability of the treated water

This work shows, for the first time, the use of the autochthonous Amazonian fruit copoazu (COPE, Theobroma grandiflorum) as an enhancer of the solar photoelectroFenton (SPEF) performance. SPEF, using a BDD anode with Na2SO4 or IrO2 anode in NaCl and actual sunlight, was applied to degrade four representative pharmaceuticals (acetaminophen, diclofenac, ciprofloxacin, and sulfamethoxazole) simultaneously in raw municipal wastewater (MWW). The effect of COPE addition on both pharmaceuticals degradation and keeping iron in solution at near-neutral pH is presented. In the SPEF process based on IrO2-NaCl, electrogenerated reactive chlorine species played the main degrading role. Meanwhile, in the BDD-based process, hydroxyl radical and photolysis were the main agents responsible for the elimination of pharmaceuticals. The two SPEF processes, IrO2-NaCl and BDD-Na2SO4, led to ~99 and ~49% of pondered degradation of pharmaceuticals, respectively. The addition of an extract of copoazu to the reactor equipped with the BDD anode improved the elimination, achieving ~70% of pondered removal of the pharmaceuticals. COPE acted as a natural complexing agent, favoring the catalytic cycle of iron in the system. Finally, the biodegradability (using aerobic microorganisms) of the MWW treated by the improved SPEF (with the BDD anode) was tested. It was found that the microorganisms were able to remove up to 54.6% of the organic carbon content in the sample, indicating the high biocompatibility of water resultant from the treatment by the SPEF improved with the copoazu presence.

Alternative Substrate with Cupuaçu Bark Promotes Morphophysiological Quality in Yellow Passion Fruit Seedlings

The use of agricultural residues for the production of alternative substrate is essential to reduce seedling production costs, as well as to reduce the environmental impacts resulting from the use of commercial substrates. Thus, the objective of this study was to evaluate the morphological and physiological performance of Passiflora edulis fruit seedlings as a function of alternative Theobroma grandiflorum bark substrates. The experiment was carried out in a screened nursery with sombrite of 50%, with a randomized block design with three replications and five plants per plot. The treatments consisted of the combination of ground and crushed cupuaçu bark in percentages of 20%, 40%, 60% and 80%. Liquid photosynthesis, stomatal conductance, internal CO2 concentration, transpiration, plant height, stem diameter, main root length, number of leaves, dry and fresh shoot and root weight and Dickson Quality Index were evaluated. The morphological and physiological performance of passion fruit plants was influenced by the cupuaçu bark concentration. Substrates containing 20 to 40% cupuaçu bark resulted in better seedlings, as they promoted increased net photosynthesis, gas exchange, leaf number, higher root growth and development, and larger stem diameter and shoot height. Consequently, the Dickson quality index showed excellent values, which shows an improvement in the biometric and physiological characteristics of the passion fruit seedlings produced with substrates based on cupuaçu bark.

Cupuassu from bean to bar: Sensory and hedonic characterization of a chocolate-like product

The processing of cupuassu (Theobroma grandiflorum Schum) beans after fermentation gives a chocolate-like product, the cupulate. The high amount of pulp adhered to the seeds hinders the fermentation. Consequently, it is necessary to depulp the seeds to perform the process, even though the pulp contains important substrates for the formation of flavor precursors. To verify whether the complete or partial removal of the pulp influences the sensory characteristics of the product, fermentation was performed with three pulp concentrations (0, 7.5, and 15%) and two schemes of turning for aeration of the mass: fixed (R1) and according to the temperature (R2), in a total of six experiments (0R1, 0R2, 7.5R1, 7.5R2, 15R1 and 15R2). The beans were processed to obtain cupulates, which were submitted to tests performed with consumers, to express their preference and attributes (acceptance, purchase intent, Check All That Apply – CATA), and then to tests with a trained panel, the Quantitative Descriptive Profile (QDP) to characterize the samples. Both tests showed the consumers’ perceptions that the cupulates have peculiar sensory characteristics. In the Consumer Test, through the Preference Mapping, all the samples of cupulates obtained from R1 conditions were preferred. In the penalty analysis, these same samples showed positive attributes that mask the negative attributes. Both CATA and QDP results showed that cupulate samples produced from seeds with a higher amount of pulp (15R1 and 15R2) had a higher number of positive mentions, for their fruity and floral flavors. The research also demonstrated that all samples gave the perception of an earthy taste, an important reason for consumer rejection, as well as a bad residual flavor. Thus, the results showed that the presence of the pulp in the fermentation environmental is important to the formation of flavor compounds and improving the sensory acceptance of the products.

Skin Regenerative Potential of Cupuaçu Seed Extract (Theobroma grandiflorum), a Native Fruit from the Amazon: Development of a Topical Formulation Based on Chitosan-Coated Nanocapsules.

Scarless skin regeneration is a challenge in regenerative medicine. Herein, we explore the regenerative potential of a Cupuaçu seed extract (Theobroma grandiflorum) to develop an innovative skin regeneration formulation based on chitosan-coated nanocapsules. Cupuaçu seed extract significantly stimulated cell proliferation and migration. A reparative gene expression profile could be verified following extract treatment, which included high levels of MKI67, a cellular proliferation marker, and extracellular matrix genes, such as ELN and HAS2, which code for elastin and hyaluronic acid synthase 2. Formulations with Cupuaçu seed extract successfully entrapped into nanocapsules (EE% > 94%) were developed. Uncoated or coated nanocapsules with low-molecular-weight chitosan presented unimodal size distribution with hydrodynamic diameters of 278.3 ± 5.0 nm (PDI = 0.18 ± 0.02) and 337.2 ± 2.1 nm (PDI = 0.27 ± 0.01), respectively. Both nanosystems were physically stable for at least 120 days and showed to be non-irritating to reconstructed human epidermis. Chitosan coating promoted active penetration into undamaged skin areas, which were still covered by the stratum corneum. In conclusion, the present study demonstrated for the first time the biotechnological potential of the frequently discarded Cupuaçu seed as a valuable pharmaceutical ingredient to be used in regenerative skin products.

Ultrasound Assisted Pretreatments Applied to Cupuaçu Husk (Theobroma grandiflorum) from Brazilian Legal Amazon for Biorefinery Concept

Cupuaçu husk (CH) is the waste of a common fruit from a native species of the Brazilian Legal Amazon. The current study investigated the influence of ultrasound (US) combined with aqueous, acid, alkaline, and ionic liquid (IL) pretreatments on the chemical and physical aspects of CH and the yield of chemical platforms production, 5-hydroxymethylfurfural (HMF) and furfural (FF), using IL. Scanning electron microscopy, X-ray diffraction (XRD) and Fourier transform infrared spectroscopy were used to feature the raw and pretreated biomass. The highest levels of glucose (9.90 g L-1) were observed in the liquid fraction resulting from the acid + US pretreatment followed by acid hydrolysis. The IL + US pretreatment recorded the best performance in removing lignin. Based on XRD analyses, ultrasound increased crystallinity of all pretreated samples as a result of the removal of cellulose’s amorphous fraction. However, it promoted accessibility to adopted reagents by increasing biomass exposure due to cavitation. The best yields of HMF and FF were recorded from hydrolysis of the solid fraction resulting from the acid + US (12.94%) and alkaline + US (48.84%) pretreatment, respectively. These results indicate satisfactory performance of ultrasound assisted pretreatments to the simplified and economic conversion of biomass into value-added products.

Analysis of the potential of cupuaçu husks (Theobroma grandiflorum) as raw material for the synthesis of bioproducts and energy generation

Lignocellulosic waste is a valuable source of energy, fuels, and raw materials for the chemical industry. In this study, the potential of cupuaçu husks, derivative from a typical fruit of the Brazilian Amazon, was evaluated. Therefore, biomass was submitted to acid hydrolysis and pyrolysis, and the contents of sugars, by-products, bio-oil compounds, and bio-oil theoretical energy potential were determined. X-ray diffraction, High performance liquid chromatography, Fourier-transform infrared and Gas chromatography mass spectrometry analyzes were used. The hydrolysate has considerable sugar levels (31.7%) and presence of relevant by-products including furfural, levulinic acid and hydroxymethylfurfural. Bio-oil contained a variety of phenolic compounds, and its theoretical energy potential suggested usability of cupuaçu husks for energy generation. Economic analysis showed that commercialization of bioproducts obtained by biomass hydrolysis and pyrolysis could be highly profitable. These outcomes indicate a range of applications for cupuaçu husks in biorefinery processes and potential to promote sustainability and social development.

Effect of adding Theobroma grandiflorum and Hylocereus polyrhizus pulps on the nutritional value and sensory characteristics of bread

This research aimed to evaluate the influence of adding cupuassu (Theobroma grandiflorum) and pitaya (Hylocereus polyrhizus) pulps on the physicochemical and sensory properties of milk bread. We prepared four formulations: control (no fruit added), and three formulations where milk was partially replaced for different proportions of pulps. The following percentages were used: cupuassu pulp (32.37 to 41.61%), pitaya pulp (4.62 to 13.87%), and milk (5.60 to 31.52%). The nutritional composition, instrumental color analysis, and acceptability of the bread were assessed. The addition of fruit pulp into the bread reduced up to 41.44% the lipid content, 40.20% of proteins, and 10.27% of caloric value. The colorimetric analysis showed that the increase in pitaya pulp and decrease in cupuassu pulp reduced the lightness (L*) and the coordinate b*, and increased the coordinate a*. The principal components analysis showed that the most accepted bread formulations were those with 18.14% and 20.74% cupuassu pulp, and 7.78% and 5.18% pitaya pulp. Thus, cupuassu and pitaya pulp can be added into bread formulations as alternative to add nutritional value and contribute to the innovation of bakery products.

Theobroma grandiflorum breeding optimization based on repeatability, stability and adaptability information

The cultivation of Theobroma grandiflorum in the Brazilian Amazon is mainly conducted by family farmers who use a range of different management strategies. Thus, breeding programs of the species must address the challenge of developing cultivars that are adapted and stable in a variety of cultivation environments. In this context, this study aimed to estimate the optimum number of harvests for genetic selection of T. grandiflorum progenies and identify the most promising ones in terms of productivity, stability, and adaptability. The trials were implemented in three environments, using a randomized complete block design, with 25 full-sib progenies, five replications, and three plants per plot. The traits mean number of fruits/plant, mean fruit production/plant, and rate of infection with witches’ broom (Moniliophthora perniciosa) were evaluated over 11 harvests. The Restricted Maximum Likelihood/Best Linear Unbiased Prediction (REML/BLUP) procedure was used to estimate genetic parameters and predict genetic values, which were then applied to assess stability and adaptability. The results show that there is genetic variability among the studied T. grandiflorum progenies and that accurate genetic selection aiming at recombination and recommendation is effective after three and eleven harvests, respectively. Six progenies were selected that met the requirements for productivity, stability, and adaptability to different cultivation environments. These results can be used to optimize and advance T. grandiflorum breeding programs.

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). β-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.

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.

Kinetic analysis of the fermentative process of mead with the addition of cupuaçu (Theobroma grandiflorum) pulp

Mead is historically one of the oldest drinks in the world. Its alcohol content can range from 4 to 14% and its formulation is basically honey, water, yeast and mineral salts, though other ingredients, such as fruits, can be added, thus giving rise to another version of mead known as melomel. Research has shown that fruity drinks have good consumer acceptability due to their alcohol content and their characteristic sweetness. Thus, the study aimed to produce mead with the addition of cupuaçu pulp and two different strains of Saccharomyces cerevisiae and Saccharomyces bayanus (T-58 and Red Star respectively), in order to perform a kinetic analysis of the fermentative process. For the elaboration of the mead, honey from the western honey bee (Apis melífera), cupuaçu pulp and yeasts were used, and the fermentation process lasted 15 days. The product was characterized through physicochemical and microbiological analyses. These analyses showed that the product is within the standards required by current Brazilian legislation. The monitoring of the kinetics of the mead fermentation allowed us to determine that the optimal time for this process is around 12 days, which is the point where there is a stabilization of the two strains in relation to the consumption of soluble solids and an increase in alcohol content. The two strains seem to be suitable for the production of mead; however, the results produced differ in alcohol content, resulting in drinks with distinct organoleptic characteristics.

Processing of fermented milk drink with different whey concentrations and addition of fruit pulps

Using whey as a complement in food formulation aiming at its utilization is an alternative to obtain new products. The present study aimed to develop a fermented milk drink with different concentrations of whey and addition of cupuaçu, soursop and açaí pulps, aiming at innovating and increasing the possibilities of production and consumption of dairy beverages. The experiment was conducted at the Food Agroindustry of the Federal Institute of Education, Science and Technology of Rondonia, Colorado do Oeste Campus, in the municipality of Colorado do Oeste, RO, Brazil. The experimental design used was completely randomized, in a 3x3 factorial scheme, consisting of three concentrations of whey (30%, 40% and 50%) and the incorporation of three fruit pulps (cupuaçu (Theobroma grandiflorum), soursop (Annona muricata) and açaí (Euterpe oleracea)), with 4 replicates. The results allowed us to conclude that whey concentration of 50% with addition of cupuaçu, soursop and açaí pulps is ideal for the production of fermented milk drink, as it leads to increased acidity and reduced values of °Brix and color parameters L and b*, making it possible to avoid the disposal of a highly nutritious compound into the environment. Whey concentration of 30% in the presence of açaí pulp enables the processing of milk drink with lower value of acidity and higher values of °Brix and color parameters L and b*. Whey concentrations of 30% and 40% in the fermented milk drinks of cupuaçu, soursop and açaí reduced acidity and increased °Brix, which suggests the possibility of using this by-product in the food industry to prepare sweeter milk drinks, meeting the minimum quality requirements according to the current legislation. The processing of whey-based dairy drinks in the presence of pulps of fruits, such as cupuaçu, soursop and açaí, constitutes an opportunity for diversification of production, improvement in the development of regional products and guarantee of food and nutritional security, respecting microbiological quality standards.

Gardnerella vaginalis and Neisseria gonorrhoeae Are Effectively Inhibited by Lactobacilli with Probiotic Properties Isolated from Brazilian Cupuaçu (Theobroma grandiflorum) Fruit.

In recent years, certain Lactobacillus sp. have emerged in health care as an alternative therapy for various diseases. Based on this, this study is aimed at evaluating in vitro the potential probiotics of five lactobacilli strains isolated from pulp of cupuaçu fruit fermentation against Gardnerella vaginalis and Neisseria gonorrhoeae. Our lactobacilli strains were classified as safe for use in humans, and they were tolerant to heat and pH. Our strains were biofilm producers, while hydrophobicity and autoaggregation varied from 13% to 86% and 13% to 25%, respectively. The coaggregation of lactobacilli used in this study with G. vaginalis and N. gonorrhoeae ranged from 15% to 36% and 32% to 52%, respectively. Antimicrobial activity was present in all tested Lactobacillus strains against both pathogens, and the growth of pathogens in coculture was reduced by the presence of our lactobacilli. Also, all tested lactobacilli reduced the pH of the culture, even in incubation with pathogens after 24 hours. The cell-free culture supernatants (CFCS) of all five lactobacilli demonstrated activity against the two pathogens with a halo presence and CFCS characterization assay together with gas chromatography revealed that lactic acid was the most abundant organic acid in the samples (50% to 62%). Our results demonstrated that the organic acid production profile is strain-specific. This study revealed that cupuaçu is a promising source of microorganisms with probiotic properties against genital pathogens. We demonstrated by in vitro tests that our Lactobacillus strains have probiotic properties. However, the absence of in vivo tests is a limitation of our work due to the need to evaluate the interaction of our lactobacilli with pathogens in the vaginal mucosa. We believe that these findings may be useful in developing a product containing our lactobacilli and their supernatants in order to support with vaginal health.

Genome Characterization of Lactiplantibacillus plantarum Strain UTNGt2 Originated from Theobroma grandiflorum (White Cacao) of Ecuadorian Amazon: Antimicrobial Peptides from Safety to Potential Applications.

The genome characterization of the Lactiplantibacillus plantarum strain UTNGt2, isolated from wild copoazu or white cacao (Theobroma grandiflorum), is described. A total of 31 contigs is assembled with a total length of 3,264,448 bases, with all contigs matching the core genome of different groups in the database. The genome size is 3,540,752 bases with GC content of 44.53% and the genome repeat sequences constitute around 457,386 bases of the assembly. The UTNGt2 matches the Lactiplantibacillus plantarum genome with 99% identity. The genome contains 3115 genes, 3052 protein-coding genes, assigned with the EggNOG database. On the basis of the results, 745 proteins are classified with an unknown function, from which 128 proteins have no match in the BLASTN database. It also contains 57 tRNAs, 5 copies of 5S rRNA, and 1 copy of tmRNA. Based on gene prediction and annotation results, 9.4% of proteins are involved in carbohydrate transport and metabolism and 8.46% in transcription, 2.36% are responsible for defense mechanisms, 0.5% are responsible for the biosynthesis of secondary metabolites, transport, and catabolism, while 25.11% have an unknown function. The genome revealed the presence of genes involved in riboflavin and folate production, the presence of CRISPR/Cas genes, phage sequences, the absence of acquired antibiotics resistance genes, virulence, and pathogenic factors, suggesting that UTNGt2 is a safe strain. Its highly antimicrobial capacity is related to the presence of two bacteriocin clusters (class IIc) of the sactipeptide class (contig 4) and plantaricin E class (contig 22), as detected by the BAGEL 4 webserver. Several RiPP-like peptides (non-bactericidal ribosomally produced and post-translationally modified peptides), polyketides (PKs), and terpenes were predicted. Whole-genome sequencing analysis revealed that the UTNGt2 strain has diverse bacteriocins with a high inhibitory capacity, thus it is a bacteriocinogenic strain. Considering the safety profile, UTNGt2 is a nonpathogenic, nonvirulent strain with valuable biotechnological traits and can be further exploited for its probiotic and antimicrobial potential in the food industry or as a potential producer strain of antimicrobial peptides as an alternative to conventional antibiotics.

Cacao, copoazu and macambo: Exploring Theobroma diversity in smallholder agroforestry systems of the Peruvian Amazon

Over the past decades, the general trend towards shade reduction and intensification of cacao management has led to biodiversity losses. In the Peruvian Amazon, the regional government is heavily promoting crop conversion to shift from regionally marketed foods towards cacao (Theobroma cacao) and copoazu (Theobroma grandiflorum). While this shift is already visibly impacting the farming landscape and the lives of many smallholder farmers, little is known about the reasons that drive farmers to choose certain types of Theobroma species or cacao varieties over others. In this paper, we addressed how cacao farmers perceive and manage specific and varietal Theobroma diversity. We interviewed cacao farmers (n = 20) during a seed-exchange fair and adapted a version of the four-square analysis to explore which Theobromas are currently adopted by farmers and why. The native cacao variety (cacao chuncho) was the one cultivated by most farmers, followed by the more industrial clonal varieties. The source of seeds and seedlings for the most cultivated varieties was a mix of donations by public institutions (for clonal varieties) and informal exchange among farmers (mainly for native and criollo varieties, and species i.e. macambo (Theobroma bicolor) and cacauillo (Theobroma speciosum)). The cacao varieties incentivized by public institutions were the least desired for future investment. The motivations for farmers to plant Theobromas was mainly based on the perceived current and potential market, but their desire to invest in a given species or variety was also based on pest resistance, traditional use, farm diversification and taste. Copoazu and CCN cacao varieties are produced by many households in large areas, while macambo, cacauillo and the cacao varieties TSH and porcelana are produced by few households in small areas. Our study suggests that agroforestry systems that include alternative Theobroma species are multi-strata and more diverse than cacao-based systems. It highlights the risks of agrobiodiversity loss associated with the promotion of industrial cacao varieties, and the importance of seed and seedling access for the development of diverse farming systems. We recommend the expansion of local seed-sharing networks and the extension of public cacao-donation campaigns to neglected and underutilized Theobromas with socio-economic and environmental benefits, in order to make the local farming systems more diverse and resilient.