Theobroma cacaoL Research Articles

Significant potential of tape yeast and plant leaves as a cover for fermented cocoa (Theobroma cacao L.) beans to generate qualities of dry cocoa beans

The purpose of this study was to determine the response to the addition of tape yeast and various cover leaves in the fermentation process of quality cocoa beans (Theobroma cacao L.) according to the Indonesian National Standard (SNI). This research was conducted at PT Perkebunan Nusantara (PTPN) XII, Ngrangkah Pawon plantation, Kediri Regency and Laboratory of Agrotechnology, Universitas Darussalam Gontor in April-August 2020. The research design used a factorial completely randomized design (CRD). The first factor was the addition of tape yeast (R) which consisted of 5 levels, namely: no yeast (R0), the addition of 0.5% tape yeast (R1), 1.0% (R2), 1.5% (R3), and 2.0% (R4). The second factor was plant leaves (D) which consisted of 5 levels, namely: D1 (banana leaves), D2 (teak leaves), D3 (taro leaves), D4 (papaya leaves) and D5 (bamboo leaves). Observation parameters included fermentation temperature of cocoa beans, pulp acidity (pH) of cocoa beans, the account of cocoa beans per 100 g, moisture content, sugar content, cut test and, phytochemical qualitative test. Observation data were analyzed by using analysis of variance (ANOVA) and further tested with LSD 5%. The addition of tape yeast and plant leaves as a cover for fermented cocoa beans improved the quality of the dry cocoa beans. There was no interaction between the addition of tape yeast and plant leaves as a cover for fermented cocoa beans in all observations of the quality of dry cocoa beans. The addition of 1% tape yeast improved the quality of fermented cocoa beans on the parameters of fermentation temperature on day 2 (43.2℃), the account of cocoa beans per 100 g (93.8 cocoa beans), sugar content (2.8°Bx) and brown seed color (99.2%). Teak leaves, taro leaves, papaya leaves and bamboo leaves can be used as a cover for fermenting cocoa beans such as banana leaves which are commonly used by farmers.

Concentraciones de glifosato y 2,4-diclorofenoxiacético en granos, hojas y suelos en cultivos de cacao (Theobroma cacao L.) en la región San Martín, Perú

El objetivo de la investigación se centró en identificar parcelas cacaoteras con presencia de glifosato y 2,4-diclorofenoxiacético; así como cuantificar estas moléculas en suelos, hojas y granos fermentados secos de cacao en 16 zonas productoras de cacao (Theobroma cacao L.) en la región San Martín. Las muestras se analizaron mediante cromatografía líquida de ultra alta resolución (UHPLC). Encontrándose concentraciones de glifosato en suelos entre 0,1751 y 0,5925 ppm; por su parte en hojas de cacao se encontraron concentraciones entre 0,0015 y 0,2639 ppm. En granos secos fermentados de cacao se obtuvieron valores de glifosato entre 0,0427 y 0,5518 ppm. En lo referente a valores de 2,4-diclorofenoxiacético, los suelos presentaron concentraciones desde 0,2248 hasta 0,7201 ppm; mientras que en hojas se reportaron valores entre 0,0011 y 0,3307 ppm. En granos secos de cacao la cuantificación fue desde 0,0708 hasta 0,6491 ppm. Se estableció que existen contrastes significativos (p < 0,05) entre las cantidades de glifosato y 2,4-diclorofenoxiacético en suelos, hojas y granos secos de cacao. Afinando que los valores de glifosato y 2,4-diclorofenoxiacético encontrados en granos secos de cacao en el 93,75% de las parcelas superan los LMR que dicta la NTS No 128-2016/MINSA/DIGESA que es de 0,1 ppm para dichos analitos.

Agronomic performance of seedlings of six cocoa(Theobroma cacaoL.) clones in the North of La Paz-Bolivia

El problema se centró en ver el comportamiento de 6 clones de cacao con buen desempeño en la Estación Experimental de Sapecho – UMSA, en otras locaciones. El objetivo es evaluar el comportamiento agronómico de seis clones de cacao con alto potencial productivo en las localidades de Tumupasa, Ixiamas y Esmeralda. La metodología consistió en trabajar con 6 tipos de clones, en parcelas con 625 plantas/ha por localidad (clon ICS 95; 104 plantas, clon III 06; 104 plantas, clon IMC 67; 104 plantas, clon ICS 06; 104 plantas, clon III 13; 104 plantas, clon ICS 01; 104 plantas) de las cuales se tomaron 5 plantas de muestra para la toma de datos. El lugar del estudio se realizó en las localidades Esmeralda, Tumupasa e Ixiamas de los municipios San Buenaventura e Ixiamas del Provincia Abel Iturralde del norte de La Paz, en el segundo semestre del 2022. En relación a los clones evaluados, se tuvo diferentes resultados, destacando el ICS-95 (42,07 cm de altura), IMC-67 (1,03 cm. diámetro de patrón), III-13 (0,63 cm diámetro de tallo); ICS-01 (12 hojas y 3,38 brotes), ICS-06 (19,07 cm longitud de hoja) y III-06 (0,43 cm diámetro de brotes). Como conclusión, los 6 clones seleccionados por la Estación Experimental de Sapecho han demostrado características de adaptación.

Enmiendas líquidas orgánicas en la reducción de cadmio en suelo, hojas y almendras de cacao

The objective of the study was to evaluate the effect of the application of two organic amendments (biol and liquid carbon) on the cadmium of the soil, leaves and almonds of the cocoa crop, plantation CCN-51 (Theobroma cacaoL.), five years of production, in the province of Huamalíes, Huánuco region, Peru. The research used a completely randomized block design (RCDB) with a factorial arrangement of 3 x 3 + 1 additional control. Total cadmium in the soil, leavesand cocoa beans was evaluated. The lowest content of total cadmium in the soil was determined when applying 50 L/ha of biol (0.13 ppm of cadmium) and when applying 4 L/ha of liquid carbon (0.12 ppm of cadmium); while, without the application of organic liquid amendments, the average value was 0.36 ppm of cadmium. Likewise, T5 (80 L/ha of biol + 4L/ha of liquid carbon) reduces the available cadmium content of the soil by 73.58%; while T4 decreases the cadmium content in leaves and almonds by 43.83% and 62.96%, respectively.

FETENSECA: Alternativa para mejorar la calidad sensorial del cacao (Theobroma cacao L.) cultivar CCN-51

The production of cocoa (Theobroma cacaoL.) in Ecuador represents the second most important economic item in agricultural exports, and its competitiveness in the international market is due to the sensory quality of the genotypes produced in the country, a quality that is developed in the post-harvest stage. For this reason, an alternative proposed by the company Agrozhucay, which according to its experience in the industry has produced the FETENSECAmethod, has been evaluated. The objective of the study was to evaluate the method of post-harvesthandling, carried out by the company AGROZHUCAY to improve the quality of cocoa in the canton of La Troncal. The methodology used was a completely randomized block design, 4 treatments with 3 replications, Duncan's multiple range test 5%, significance level equal to 95%. Physical properties and biomolecules were studied using the DPPH and Folin Ciocalteu methods. The results determine that the T3 and T0 treatment presented the best physical-chemical characteristics of the best treatments, recommending these treatments to improve the sensory and physical-chemical quality

Plant metal concentrations in Theobroma cacao as affected by soil metal availability in different soil types

Beans of cacao (Theobroma cacaoL.) are used to produce a variety of chocolate products. Bioaccumulation of metals at toxic levels through the consumption of contaminated products has been identified as a health concern in humans. Both metal diversity and concentration as well as their interactions in the soil influence essential and non-essential metal uptake in plants; but the effects of these on bioaccumulation of metals in cacao is not understood across diverse soil types. In this study eight metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) were investigated in 12 soil subgroups belonging to four soil orders across 15 locations in Trinidad, with the aim to investigate the effect of soil metal diversity and concentration on metal bioaccumulation in cacao. Soil metals were extracted using five methods (aqua regia, DTPA, Mehlich 3, nitric acid, and water). Cacao leaf metal concentrations were determined using the USEPA 3052 method. Metal extraction efficiency ranged between methods with aqua regia ≥ nitric acid > Mehlich 3 ≥ DTPA ≥ water across all metals. The soil extraction method that best predicted cacao leaf metal concentrations varied with the metal – Mehlich 3 or DTPA for Cd, Ni, Zn; aqua regia, Mehlich 3, or nitric acid for Pb, and water for Mn. A stepwise regression analysis showed that plant metal concentration can be predicted using soil physicochemical characteristics as well as the concentration of metals in the soil. The importance of soil type on cacao leaf metal bioaccumulation is discussed.

Green simple microwave-assisted extraction (MAE) of cellulose from Theobroma cacao L. (TCL) husk

This work had successfully produced Theobroma cacao L. (TCL) husk-based cellulose used efficiently and friendly environmentally simple microwave-assisted extraction (MAE) method. The microwave-assisted heating time optimization of alkaline-treatment was carried out for 10, 20, 30 and 40 minutes. Microwave-assisted bleaching process was also conducted using green bleaching agent H2O2, and also utilizing microwave heating for 60 minutes. The followed treatment was filtered, neutralized, washed and overnight freeze-dried. Fourier transform infrared (FTIR) analysis confirmed common cellulose functional groups of TLC, are β-(1,4)-glycosidic bonds at ∼897 cm-1, O-H (hydrogen bond) at 3412 cm-1, C-H vibration at 2902 cm-1, C-O-C asymmetric at 1161 cm-1, and C-OH out-of-plane bending at 665 cm-1. FTIR analysis, in addition, also studied crystallinity ratio (CrR), hydrogen bond energy (EH) and hydrogen bond distances (RH), while particle size analyzer (PSA), X-ray diffraction (XRD) and thermogravimetry analysis (TGA) were carried out to confirm particles sizes, crystallinities and thermal stability properties of TCL husk, cellulose of TCL husk and commercial-microcrystalline cellulose (MCC), respectively.

Yield Performance of the Promising Cocoa Hybrids (Theobroma cacaoL.) at Dry Climate Condition

Global climate change is impacting cocoa cultivation that cocoa trees must be adaptive in dry climate and higher temperature of environmental condition. This research had objective to observe adaptability performance of the promising hybrids of cocoa ofthe selecting high yielding hybrid swhichare tolerantto dry condition. Treatments were 14 hybrids of bi-clonal crosses betweenselected clones in factorial design in which KEE 2 was selected based on its resistance to vascular-streak dieback (VSD) and drought condition. Multilocation trials were established in Kaliwining Experimental Station in Jember (low land) and Sumber Asin Experimental Station in Malang (medium land) both of which were classified to dry land area. Yield per tree were assessed in monthly basis during 9 consecutive years of yield period. Data were performed to the combinedanalysis of variance followed by AMMI (additive main effect and multiplicative interaction). The results showed thatthe factor of hybrid, location, and interactionof hybrid and location to year contributed higher value of variance for the yield. Analysis of AMMI indicated that the hybrids of KEE 2 x Sulawesi 1 (F 1 and reciprocal) had highest yield potency and adaptive to low land (Kaliwining) on the other hand the hybrids of TSH 858 x KEE 2 (F 1 and reciprocal) were adaptive to medium land (Sumber Asin). Refer to yield adaptability to dry condition and other potential traits reported thus the hybrid of KEE 2 x Sulawesi 1 (F 1 and reciprocal) was released as cocoa planting material for addressing dry condition. The result also identified those hybrids inherited using KEE 2 performed more adaptive to dry condition. Further-more, the clone was suggested to be used as parental clone on breeding for drought tolerance.

Role of Exogenous Salicylic Acid and Benzoic Acid Applications to Vascular Streak Dieback Disease Attack on Cocoa Seedlings

Currently, vascular streak dieback (VSD) attacks have widespread in areas ofthe center of cocoa (Theobroma cacaoL.) production, especially inSulawesi. Salicylic acid and benzoic acid are part of phenol compounds reportedlyacts as bioimmunoregulator in increasing of plant resistance to that pathogen. The objec-tiveof this research was to investigate effect of exogenous salicylic acid and benzoic acid applications to VSD disease attack on cocoa seedlings. The research was conducted at Indonesian Coffee and Cocoa Research Institute,Jember, East Java. This experiment designed using randomized completely block design with three replications. The treatments included concentrations of salicylic acid in 0.5 mMol, 1 mMol, 2 mMol; benzoic acid with concentration 0.5 mMol, 1 mMol,2 mMol; and control (water spray). Observations were based on the incidence and severity damage of disease, plant height, stem diameter, number of leaves, stelate trichome, chlorophyll, width and length of stomatal apperture, and number of stomata. Exogenoussalicylic acid and benzoic acid as phytohormone can reduce disease insidence and severity of VSD. In conclusion, our results indicated that exogenous salicylic acid and benzoic promoted stomatal closure in cocoa leaf by decreasing width-stomatal closure. Salicylic and benzoic acid had no effect to plant height, stem diameter, number of leaves, length of stomatal apperture, number of stomata, and number of stelate trichome.

Heterosis Analysis of Leaf Stomatal Characteristics on F 1 Population of Cocoa (Theobroma cacao L.) Related to Vascular-Streak Dieback Resistance

Breeding for cocoa resistance to VSD is the main goal in breeding program in Indonesia. Research for selection criteria on cocoa (Theobroma cacaoL.) resistance to vascular-streak dieback (VSD) has being carried out by exploring leaf stomatal characteristics corresponding to VSD damages. This research had objectives to select best crosses by observing the estimated value of heterosis and heterobeltiosis on F1 hybrid populations based on leaf stomata characteristics related to VSD resistance. Trial was established at Kaliwining Experimental Station of Indonesian Coffee and Cocoa Research Institute (ICCRI) in a VSD endemic area. Treatments consisted of 5 F1 hybrid populations generated by biparental crosses of TSH 858(S) x Sulawesi 1 (R), TSH 858 (S) x KEE 2 (R), TSH 858 (S) x Sulawesi 3 (R), Sca 6 (R) x KW 264 (S) and KW 617 (R) x KW 264 (S). Each cross consisted of 100-300 genotypes planted in high planting density of 0.5 m x 0.5 m. Theresistance was evaluated by scoring the plant damage in the scale of 0-3 at three years after planting. Stomata characteristics were assessed microscopically based on the variables of density, length, width and diameter of stomata and width of opening stomata. Analysis of correlation indicated that most of stomatal charac-teristicssignificantly correlated to VSD damage but the width of opening stomata performedthe highest coefficient of correlation (r = 0.49) and contributed to VSD damage (0.45). Meanwhile, path analysis indicated higher value of the char-acteristics performed less resistance of the tested hybrids, KW 617 x KW 264 showed lower value of heterosis and heterobeltiosis for the characteristics which of the best crosses for generating VSD-resistant genotype.

EFFECT OF ORGANIC AND INORGANIC AMENDMENTS ON PRODUCTIVITY OF COCOA ON A MARGINAL SOIL IN SULAWESI, INDONESIA

SUMMARYReduced soil fertility and damage from pests and diseases have contributed to a decline in productivity of cocoa (Theobroma cacaoL.) smallholdings in Sulawesi, Indonesia over the last decade. In a trial on a marginal, acidic soil in South Sulawesi, young PBC123 cocoa trees were supplied with compost, mineral fertiliser (NPK fertiliser and urea) or dolomite, alone and in combination. After 20 months, the trees supplied with compost were taller, flowered more profusely and had a five-fold higher dry bean yield than other treatments. Treatments had no impact on incidence of cocoa pod borer, Phytophthora pod rot and vascular streak dieback. All of the trees supplied with compost survived, while the control, mineral-fertiliser- and dolomite-treated trees had a mortality rate of 22–45% and symptoms of interveinal necrosis. Leaf concentrations of N, P and K were within the normal range in all treatments. In the control and mineral fertiliser treatments, leaf concentrations of Ca (0.28–0.30%) and Mg (0.11–0.15%) were deficient, but were higher in trees supplied with compost (0.78–1.21% and 0.26–0.29%, respectively). The Mg/K ratio in soil-exchangeable cations and leaves was increased three-fold by the combined compost/dolomite treatment. Supplying mineral fertiliser alone resulted in 3.3 cmol kg−1exchangeable Al, compared to 2.2 cmol kg−1in control soils. Since 10 kg tree−1year−1compost was supplied, a rate that is not practical on most cocoa smallholder farms, further investigation of cost-effective applications of organic matter in conjunction with appropriate formulations of inorganic fertilisers is recommended.

STUDY OF TREE-TO-TREE YIELD VARIABILITY AMONG SEEDLING-BASED CACAO POPULATIONS IN AN INDUSTRIAL PLANTATION IN CÔTE D'IVOIRE

SUMMARYIn Côte d'Ivoire, the world's largest cocoa producer, cacao (Theobroma cacaoL.) is usually grown from seed. The genetics consist of a mix of amelonado, trinitario and selected hybrids. This mix of varieties generates high phenotypic variabilities, including variability in tree productivity, within cacao populations in both smallholder and industrial plantations. Tree-to-tree variability in yield has been reported in cacao breeding trials under different environmental conditions. However, it has never been considered a limiting factor for agronomical performance of seedling-based cacao plantations. Around 10 000 cacao trees from seven plots under different environmental conditions in a cacao plantation in Côte d'Ivoire were monitored for 2 years. Pod production of individual trees was recorded and annual average tree pod yields were assessed. High heterogeneity in cacao-tree yields was observed in all plots, with coefficients of variation ranging from 56 to 102%. The distribution of cacao-tree yields in each plot was positively skewed. Analysis of these distributions showed that unproductive trees represented a significant proportion of cacao tree populations (7%), and the 20% least productive trees accounted for 3% of production. The 20% most productive trees were responsible for 46% of the total pod production of a plot. This heterogeneity reflects a major imbalance in the agronomical performances of low- and high-yielding trees and also represents possible efficiency gaps in seedling-based cacao plantations, which could be overcome through innovative corrective strategies, opening new pathways for improvement of cacao-based cropping systems.

COCOA AND TOTAL SYSTEM YIELDS OF ORGANIC AND CONVENTIONAL AGROFORESTRY VS. MONOCULTURE SYSTEMS IN A LONG-TERM FIELD TRIAL IN BOLIVIA

SUMMARYCocoa (Theobroma cacaoL.) is produced in systems of varying complexity ranging from monoculture with temporary shade to highly diverse agroforests. Producers have to weigh high cocoa yields in the short to medium term in monocultures against higher total system yields in the short term and sustainable production systems in the long term in conjunction with ecosystem services in agroforestry systems (AFs). More long-term data on the comparative agronomic, economic and ecological performance of differently managed cocoa production systems is required to make sound recommendations to farmers. This paper describes the only long-term field trial worldwide comparing different cocoa production systems under conventional and organic management. The factors (i) crop diversity (monoculture vs. agroforestry), (ii) management practice (conventional vs. organic) and (iii) cultivar are being tested in a full-factorial, randomized complete block design with four replications. First, results showed significantly faster development of trunk circumferences in monocultures (+21%) compared to AFs. Cocoa yields were 47% lower in the organic compared to the conventional monoculture. In the AFs, however, the organic–conventional yield gap was less pronounced (−16%) and statistically insignificant. The cumulative yields of all products harvested were significantly higher in the AFs (+161%) compared to the monocultures. The productivity of cocoa by-crops in AFs may contribute to local food security and risk distribution in smallholder contexts.

Efek Temperatur Terhadapvirulensi Jamur Colletotrichum Gloeosporioidespenz. Sacc.penyebab Penyakit Antraknosa Padatanaman Kakao(theobroma Cacao L.)

Temperature Effect TowardsVirulency ofColletotrichum gloeosporioides Penz. Sacc. FungalDisesase Caused of Anthracnose on Cocoa(Theobroma cacaoL.).This research aims to determindvirulency level, disease incident and temperature effect towards fungal morphology growth of C.gloeosporioides fungal disesase caused of anthracnose oncocoa (Theobroma cacao L.). The studywas conductedat Karang Anyar village, regency in Langkat with altitude±28masl, at Sambirejovillage, regency in Langkat with altitude±28masl, at Bekukul village, regency in Deli Serdang withaltitude±100 masl, at Sayum Sabah village, regency in Deli Serdang with altitude±200 masl and atthe Laboratory ofPlant Disease, Agroecotechnology Program Study, Faculty of AgricultureUniversity of Sumatera Utara, Medan from July to December 2012.The method of this research issurvey.Results showedthat highest attack percentage was founded in Sambirejo villagewhich reach100% and thelowest was founded in Sayum Sabah village which reach 95%, highest diseaseseverity was founded in Bekukul village which reach 92.0% and thelowest was founded in KarangAnyar village which reach 78.6% and showed no differences fungus morphology that extractedfrom four different location.Keywords: C. gloeosporioides,virulency, morphology, temperature, cocoa

Development of Selection Criteria on Bean Weight Character of Cocoa (Theobroma cacaoL.) through Path Analysis Approach

Path coefficient analysis is frequently used for development of selection criteria on various type of plants. Path analysis on this research was conducted to find the selection criteria of yield component which directly affect bean weight. In addition to the value of path analysis coefficient, genetic variation coefficient, heritability and the value of genetic progress were also studied. The study was conducted at the Indonesian Coffee and Cocoa Research Institute. The research used randomized complete block design consisting of 14 accession numbers and each consisting of three replications. Pod girth, pod length, pod weight, wet beans weight per pod, number of normal beans per pod, number of abnormal beans per pod, dry weight per normal bean, and shell content were observed. The results showed that the pod weight character had an important role in determining the dry weight of normal bean. The character had a positive genotype correlation coefficient values which was high and significantly different (r=0.46) for dry weight per normal bean, considerable direct influence (P=0.479), moderate of the genotype variation coefficient (9.6%), and high genetic progress (95.23). Character of wet bean weight per pod could also be used indirectly for the selection criteria for dry weight per normal bean based on genetic variation coefficient value (11.88%), genetic progress value (82.48), and direct effect on dry weight per normal bean had positive value (P=0.006). Key words: Selection criteria, dry weight per bean, path analysis,Theobroma cacaoL.

Two fruits per pedicel in cacao (Theobroma cacaoL.) – a rare phenotype

A rare aberrant fruit phenotype was found in cacao (Theobroma cacaoL.) in St Augustine, Trinidad. Double fructification instead of single fructification on a single pedicel was observed. The aberration probably occurred as a result of the formation of two pistils in a single flower. Fruits matured as normal, and fruit morphology and seed number were within the range of that reported for normal fruits of the accession observed in Trinidad. The plant is under investigation and the impact of this finding is highlighted.

카카오(Theobroma cacao L.) 용매 분획물의 항산화 및 항염증 효과

본 연구는 카카오를 화장품 소재로 활용하기 위하여 항산화 및 항염증 효과를 검증하여 기능성 화장품 소재로서의 가능성을 검토하였다. 아세톤 추출물을 이용하여 클로로포름층, 에틸아세테이트층, 부탄올층, 물층의 극성별로 분획을 실시하였다. 카카오 분획물의 전자공여능은 100 <TEX>${\mu}g/ml$</TEX>에서 에틸아세테이트 분획물이 76.2%, 부탄올 분획물이 53.9%의 소거능을 나타내었다. 또한, superoxide anion radical 소거능 측정 결과, 에틸아세테이트 분획물과 부탄올 분획물이 50 <TEX>${\mu}g/ml$</TEX>에서 각각 76.09%, 51.4%의 소거능이 나타났고, <TEX>$Fe^{2+}$</TEX>, <TEX>$Cu^{2+}$</TEX>의 첨가에 따른 지방산패 억제능에서 <TEX>$Fe^{2+}$</TEX>를 첨가한 군에서는 에틸아세테이트 분획물이 50 <TEX>${\mu}g/ml$</TEX>에서 64%로 <TEX>$Cu^{2+}$</TEX>를 첨가한 군보다 저해능이 높았다. 항염증 효과 측정으로 hyaluronidase 저해 활성을 측정한 결과, 부탄올 분획물의 경우 100 <TEX>${\mu}g/ml$</TEX>에서 53.04%의 저해활성을 나타내었으며, lipoxygenase 저해 활성 측정결과 10 <TEX>${\mu}g/ml$</TEX>에서 51.32%의 저해활성을 나타내었다. 대식세포를 이용한 nitric oxide (NO) 저해활성을 측정한 결과 카카오 에틸아세테이트 분획물에서 가장 높은 NO 저해활성을 나타냄을 확인할 수 있었다. 또한 대식세포 내에서 에틸아세테이트 분획물 100 <TEX>${\mu}g/ml$</TEX> 농도에서 iNOS, COX-2 단백질 발현을 저해하였다. Solvent extracts of Theobroma cacao L. (TCL) were investigated for anti-oxidative and anti-inflammatory effects in order to consider TCL as a functional ingredient for cosmetic products. TCL(A) extract was fractioned according to polarity with <TEX>$CHCl_3$</TEX>, EtOAc, n-BuOH, and water. Following TCL(A) fractionation, the electron-donating ability of the n-BuOH and EtOAc solvent fractions (each 100 <TEX>${\mu}g/ml$</TEX>) was about 76.2% and 53.9%, respectively. The superoxide anion radical inhibitory effect of the n-BuOH and EtOAc solvent fractions (each 50 <TEX>${\mu}g/ml$</TEX>) was about 76.09% and 51.4%, respectively. Results of lipid oxidation showed that <TEX>$Fe^{2+}$</TEX> had a greater chelating effect than <TEX>$Cu^{2+}$</TEX>. The <TEX>$Fe^{2+}$</TEX> chelating effect of the EtOAc solvent fraction (50 <TEX>${\mu}g/ml$</TEX>) was about 64%. Hyaluronidase inhibition related to the anti-inflammatory effect was 53.0% with EtOAc at 100 <TEX>${\mu}g/ml$</TEX>, while the lipoxygenase inhibitory effect was about 51.32% at 10 <TEX>${\mu}g/ml$</TEX>. The anti-inflammatory activity in the EtOAc fraction inhibited the generation of nitric oxide. Results also showed that iNOS protein expression increased in RAW264.7 cells. In contrast, at 100 <TEX>${\mu}g/ml$</TEX> EtOAc, iNOS and COX-2 protein expression significantly decreased in LPS-stimulated RAW 264.7 cells.

Scientific Opinion on the substantiation of health claims related to caffeine and theobromine in cocoa (Theobroma cacaoL.) and enhancement of mood (ID 4276) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

Scientific Opinion on the substantiation of health claims related to caffeine and theobromine in cocoa (Theobroma cacaoL.) and enhancement of mood (ID 4276) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

Microsatellite fingerprinting in the International Cocoa Genebank, Trinidad: accession and plot homogeneity information for germplasm management

The International Cocoa Genebank, Trinidad, is the largest field genebank collection of cacao (Theobroma cacaoL.) in the public domain and the correct identity of each tree is crucial for germplasm movement, evaluation and phenotypic characterization. Nine microsatellite loci were used to assess the identity of 1477 trees from 486 cacao accessions representing approximately 16.9% of the trees and 29.2% of the accessions within the genebank. Heterogeneous plots (plots containing more than one genotype group) averaged 25.1% in The International Cocoa Genebank, Trinidad, with maximal admixture (32.6%) being recorded in Field 5B. The error rate did not differ significantly among different fields. Mislabeling error could be affected by accession grouping with an average error rate of 27.4% for accession groups in the genebank. Synonymous accessions were estimated to account for 14.4% of the field genebank. The results of the present study provide essential information for the management and utilization of the germplasm collection. Single-tree genotyping of every tree in this collection is strongly recommended.

The relic Criollo cacao in Belize – genetic diversity and relationship with Trinitario and other cacao clones held in the International Cocoa Genebank, Trinidad

Cacao (Theobroma cacaoL.) was domesticated in Mesoamerica and is native to the South American rainforest. Belizean Criollo is a group of relic landraces that are thought to be similar to those used by the Olmecs and Mayans during early domestication. Knowledge of genetic diversity is essential for efficient conservation and use of these relic landraces. Using 30 microsatellite markers, we characterized genetic diversity in 77 Belize Criollo accessions collected from the Maya Mountains in Belize, and assessed their relationship with 62 cacao accessions including 25 Trinitario accessions. Genetic diversity and heterozygosity were low in Belizean Criollo germplasm. Eleven distinctive genotypes were identified among the Belizean germplasm. Results of ordination and cluster analysis supported their putative ancestral contribution to the Trinitario cacao. However, results of Bayesian assignment and parentage analysis both suggested that the contribution of Criollo cacao to the Imperial College Selections Trinitario is small. Our preferred hypothesis for the genesis of Trinitario cacao is that a limited population of Criollo × Forastero hybrids emanated from the introduced Forastero population of Trinidad. The present study provides new insights into the origin of the Trinitario cacao, which will be useful in theex situandin situconservation of cacao landraces from Mesoamerica.