The genome sequence of the most widely cultivated cacao type and its use to identify candidate genes regulating pod color

USDA-ARS?s Scientific Manuscript database

Background: Theobroma cacao L. cultivar Matina 1-6 belongs to the most cultivated cacao type. The availability of its genome sequence and methods for identifying genes responsible for important cacao traits will aid cacao researchers and breeders. Results: We describe the sequencing and assembly of...

Coffee vs. Cacao: A Case Study from the Vietnamese Central Highlands

ERIC Educational Resources Information Center

Ha, Dang Thanh; Shively, Gerald

2005-01-01

Mr. Nam, the vice chair of a village in Dak Lak province of Vietnam, was keen to protect farmers in his village from the sharp decline in prices of coffee ("Coffea canephora" Pierre ex Froehner). He did this by encouraging farmers in his village to plant cacao ("Theobroma cacao" L. subsp. "cacao"). Cacao was suitable…

Molecular characterization of an earliest cacao (Theobroma cacao L.) collection from Peruvian Amazon using microsatllite DNA markers

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is indigenous to the Amazon region of South America. The Peruvian Amazon harbors a large number of diverse cacao populations. Since the 1930s, several numbers of populations have been collected from the Peruvian Amazon and maintained as ex situ germplasm repositories in ...

Macro and micro nutrient uptake parameters and use efficiency in cacao genotypes influenced by deficient to excess levels of soil K

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is an important economic crop for many of the tropical countries. Adequate levels of soil K are essential for good growth and achieving high cocoa bean yields. Soils under cacao invariably have low levels of plant available K to support good cacao growth. Growth chamber ex...

Heavy metal accumulations in cacao (Theobroma cacao L.) leaves and cocoa beans grown at three main cacao growing regions of Peru

USDA-ARS?s Scientific Manuscript database

Peru is one of the leading exporters of organic cocoa beans in the world. However, the accumulation of heavy metals in cacao beans represents a considerable quality problem. The aim of this study was to investigate the distribution and accumulation of heavy metals in cacao plants grown at three dif...

Soil physical and chemical properties of cacao farms in the south western region of cameroon

USDA-ARS?s Scientific Manuscript database

The low macro nutrient content (K, Ca and Mg) in soils under cacao is one of the major causes of the poor cacao (Theobroma cacao L) yields. Efforts were made to assess the major physical and chemical properties of soils from some important cacao zones of the South West Region of Cameroon in order t...

Accurate determination of genetic identity for a single cacao bean, using molecular markers with a nanofluidic system, ensures cocoa authenticity and traceability

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is an important tropical crop since it is the source of cocoa butter and powder for the confectionery industry. Production and marketing of premium high-value fine flavored cacao provide opportunities for cacao growers, the chocolate industry and consumers. The higher far...

Comparison of polyphenol concentration and composition between genetically diverse cacao (Theobroma cacao L.) accessions selected for high yield and disease resistance

USDA-ARS?s Scientific Manuscript database

There is abundant evidence that consumption of cacao and dark chocolate promotes human health and that the main cacao components contributing positive health effects are polyphenols. The polyphenols in cacao bean constitute 12–18% dry weight of the whole bean and are predominantly catechins (37% w/w...

Chloroplast microsatellite primers for cacao (Theobroma cacao) and other Malvaceae.

PubMed

Yang, Ji Y; Motilal, Lambert A; Dempewolf, Hannes; Maharaj, Kamaldeo; Cronk, Q C B

2011-12-01

Chloroplast microsatellites were developed in Theobroma cacao to examine the genetic diversity of cacao cultivars in Trinidad and Tobago. Nine polymorphic microsatellites were designed from the chloroplast genomes of two T. cacao accessions. These microsatellites were tested in 95 hybrid accessions from Trinidad and Tobago. An average of 2.9 alleles per locus was found. These chloroplast microsatellites, particularly the highly polymorphic pentameric repeat, were useful in assessing genetic variation in T. cacao. In addition, these markers should also prove to be useful for population genetic studies in other species of Malvaceae.

Genetic diversity and parentage in farmer varieties of cacao (Theobroma cacao L.) from Honduras and Nicaragua as revealed by Single Nucleotide Polymorphism (SNP) markers

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is the main source for chocolate with an annual production of four million tons worldwide. This Neotropical tree crop was domesticated in Mesoamerica as far back as 3,000 years ago. Knowledge of genetic diversity and population structure in farmer varieties of cacao in the...

Phytophthora megakarya and P. palmivora, closely related causal agents of cacao black pod induce similar reactions when infecting pods of a susceptible cacao genotype

USDA-ARS?s Scientific Manuscript database

Phytophthora megakarya (Pmeg) and Phytophthora palmivora (Ppal) cause black pod rot of Theobroma cacao. Of these two clade 4 species; Pmeg is more virulent and is displacing Ppal on cacao in many cacao production areas in Africa. To understand the advantages Pmeg has over Ppal, we compared symptom...

Genetic identity, ancestry and parentage in farmer selections of cacao from Aceh, Indonesia revealed by single nucleotide polymorphism (SNP) markers

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is the source of cocoa powder and butter used for chocolate and this species originated in the rainforests of South America. Indonesia is the 3rd largest cacao producer in the world with an annual cacao output of 0.55 million tons. Knowledge of on-farm genetic diversity is...

First typology of cacao (Theobroma cacao L.) systems in Colombian Amazonia, based on tree species richness, canopy structure and light availability

PubMed Central

Suárez Salazar, Juan Carlos; Melgarejo, Luz Marina; Di Rienzo, Julio A.; Casanoves, Fernando

2018-01-01

Aim and background We present a typology of cacao agroforest systems in Colombian Amazonia. These systems had yet to be described in the literature, especially their potential in terms of biodiversity conservation. The systems studied are located in a post-conflict area, and a deforestation front in Colombian Amazonia. Cacao cropping systems are of key importance in Colombia: cacao plays a prime role in post conflict resolution, as cacao is a legal crop to replace illegal crops; cacao agroforests are expected to be a sustainable practice, promoting forest-friendly land use. Material and methods We worked in 50 x 2000 m2 agroforest plots, in Colombian Amazonia. A cluster analysis was used to build a typology based on 28 variables characterised in each plot, and related to diversity, composition, spatial structure and light availability for the cacao trees. We included variables related to light availability to evaluate the amount of transmitted radiation to the cacao trees in each type, and its suitability for cacao ecophysiological development. Main results We identified 4 types of cacao agroforests based on differences concerning tree species diversity and the impact of canopy spatial structure on light availability for the cacao trees in the understorey. We found 127 tree species in the dataset, with some exclusive species in each type. We also found that 3 out of the 4 types identified displayed an erosion of tree species diversity. This reduction in shade tree species may have been linked to the desire to reduce shade, but we also found that all the types described were compatible with good ecophysiological development of the cacao trees. Main conclusions and prospects Cacao agroforest systems may actually be achieving biodiversity conservation goals in Colombian Amazonia. One challenging prospect will be to monitor and encourage the conservation of tree species diversity in cacao agroforest systems during the development of these cropping systems, as a form of forest-friendly management enhancing sustainable peace building in Colombia. PMID:29401499

Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses

PubMed Central

Abou Rajab, Yasmin; Leuschner, Christoph; Barus, Henry; Tjoa, Aiyen; Hertel, Dietrich

2016-01-01

One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha-1), total net primary production rose twofold (from 9 to 18 Mg C ha-1 yr-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1–1.2 Mg C ha-1 yr-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity. PMID:26927428

Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses.

PubMed

Abou Rajab, Yasmin; Leuschner, Christoph; Barus, Henry; Tjoa, Aiyen; Hertel, Dietrich

2016-01-01

One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha-1), total net primary production rose twofold (from 9 to 18 Mg C ha-1 yr-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1-1.2 Mg C ha-1 yr-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity.

Effects of different agricultural systems on soil quality in Northern Limón province, Costa Rica.

PubMed

Cornwell, Emma

2014-09-01

Conversion of native rainforest ecosystems in Limón Province of Costa Rica to banana and pineapple monoculture has led to reductions in biodiversity and soil quality. Agroforestry management of cacao (Theobroma cacao) is an alternative system that may maintain the agricultural livelihood of the region while more closely mimicking native ecosystems. This study compared physical, biological and chemical soil quality indicators of a cacao plantation under organic agroforestry management with banana, pineapple, and pasture systems; a native forest nearby served as a control. For bulk density and earthworm analysis, 18 samples were collected between March and April 2012 from each ecosystem paired with 18 samples from the cacao. Cacao had a lower bulk density than banana and pineapple monocultures, but greater than the forest (p < 0.05). Cacao also hosted a greater number and mass of earthworms than banana and pineapple (p < 0.05), but similar to forest and pasture. For soil chemical characteristics, three composite samples were collected in March 2012 from each agroecosystem paired with three samples from the cacao plantation. Forest and pineapple ecosystems had the lowest pH, cation exchange capacity, and exchangeable nutrient cations, while cacao had the greatest (p < 0.05). Total nutrient levels of P and N were slightly greater in banana, pineapple and pasture than in cacao; probably related to addition of chemical fertilizer and manure from cattle grazing. Forest and cacao also had greater %C, than other ecosystems, which is directly related to soil organic matter content (p < 0.0001). Overall, cacao had more favorable physical, biological and chemical soil characteristics than banana and pineapple monocultures, while trends were less conclusive compared to the pastureland. While organic cacao was inferior to native forest in some soil characteristics such as bulk density and organic carbon, its soil quality did best mimic that of the native forest. This supports the organic cultivation of cacao as a desirable alternative to banana and pineapple monoculture.

Horticultural traits associated with cacao accessions recommended for Puerto Rico

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is an important agricultural product from which the international chocolate industry is based upon. Increasing demand for chocolate, especially in emerging markets in Asia, coupled with reduced worldwide production has led to shortfalls in cacao ‘bean’ supplies. Deficits...

Better chocolate through genomics

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the cacao or chocolate tree, is a tropical understory tree whose seeds are used to make chocolate. And like any important crop, cacao is the subject of much research. On September 15, 2010, scientists publicly released a preliminary sequence of the cacao genome--which contains all o...

Evidence of cacao use in the Prehispanic American Southwest

PubMed Central

Crown, Patricia L.; Hurst, W. Jeffrey

2009-01-01

Chemical analyses of organic residues in fragments of ceramic vessels from Pueblo Bonito in Chaco Canyon, New Mexico, reveal theobromine, a biomarker for cacao. With an estimated 800 rooms, Pueblo Bonito is the largest archaeological site in Chaco Canyon and was the center of a large number of interconnected towns and villages spread over northwestern New Mexico. The cacao residues come from pieces of vessels that are likely cylinder jars, special containers occurring almost solely at Pueblo Bonito and deposited in caches at the site. This first known use of cacao drinks north of the Mexican border indicates exchange with cacao cultivators in Mesoamerica in a time frame of about A.D. 1000–1125. The association of cylinder jars and cacao beverages suggests that the Chacoan ritual involving the drinking of cacao was tied to Mesoamerican rituals incorporating cylindrical vases and cacao. The importance of Pueblo Bonito within the Chacoan world likely lies in part with the integration of Mesoamerican ritual, including critical culinary ingredients. PMID:19188605

Physicochemical properties and antioxidant capacity of raw, roasted and puffed cacao beans.

PubMed

Hu, SuJung; Kim, Byung-Yong; Baik, Moo-Yeol

2016-03-01

The antioxidant capacity and attributable bioactive compounds of puffed cacao beans were investigated. Roasting was carried out at 190°C for 15min and puffing was performed at 4-7kgf/cm(2). Cacao beans puffed at 4kgf/cm(2) showed the highest total polyphenols (23.16mgGAE/gsample) and total flavonoids (10.65mgCE/gsample) (p<0.05). As the puffing pressure increased, the amount of total polyphenols and total flavonoids decreased. The antioxidant capacity of cacao beans reflected the total polyphenols and flavonoids measured. The quantities of theobromine, catechin, epicatechin, and procyanidin B2 were higher in cacao beans puffed at 4kgf/cm(2) than in roasted cacao beans. Puffed cacao beans received a good sensory score in flavor, but sourness increased as puffing pressure increased. Thus, these results suggest that, in cacao bean processing, puffing could be an alternative to roasting, which provide a rich taste and high antioxidant capacity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Colombia a Source of Cacao Genetic Diversity As Revealed by the Population Structure Analysis of Germplasm Bank of Theobroma cacao L.

PubMed

Osorio-Guarín, Jaime A; Berdugo-Cely, Jhon; Coronado, Roberto Antonio; Zapata, Yeny Patricia; Quintero, Constanza; Gallego-Sánchez, Gerardo; Yockteng, Roxana

2017-01-01

Beans of the species Theobroma cacao L., also known as cacao, are the raw material to produce chocolate. Colombian cacao has been classified as a fine flavor cacao that represents the 5% of cacao world's production. Colombian genetic resources from this species are conserved in ex situ and in-field germplasm banks, since T. cacao has recalcitrant seeds to desication and long-term storage. Currently, the collection of T. cacao of the Colombian Corporation of Agricultural Research (CORPOICA) has approximately 700 germplasm accessions. We conducted a molecular analysis of Corpoica's cacao collection and a morphological characterization of some accessions with the goal to study its genetic diversity and population structure and, to select interesting accessions for the cacao's breeding program. Phenotypic evaluation was performed based on 18 morphological traits and 4 biochemical traits. PCA analysis of morphological traits explained 60.6% of the total variation in seven components and 100% of the total variation of biochemical traits in four components, grouping the collection in 4 clusters for both variables. We explored 565 accessions from Corpoica's germplasm and 252 accessions from reference populations using 96 single nucleotide polymorphism (SNP) molecular markers. Molecular patterns of cacao Corpoica's collection were obtained amplifying specific alleles in a Fluidigm platform that used integrated circuits of fluids. Corpoica's collection showed highest genetic diversity [Expected Heterozygosity ( H E = 0.314), Observed Heterozygosity ( H O = 0.353)] that is reduced when reference populations were included in the dataset ( H E = 0.294, H O = 0.261). The collection was divided into four clusters based on population structure analysis. Cacao accessions from distinct groups showed some taxonomic concordance and reflected their geographic origins. For instance, accessions classified as Criollo were clearly differentiated in one group and we identified two new Colombian genetic groups. Using a number of allelic variations based on 87 SNP markers and 22 different morphological/biochemical traits, a core collection with a total of 232 accessions was selected as a primary genetic resource for cacao breeders.

The genome of Theobroma cacao.

PubMed

Argout, Xavier; Salse, Jerome; Aury, Jean-Marc; Guiltinan, Mark J; Droc, Gaetan; Gouzy, Jerome; Allegre, Mathilde; Chaparro, Cristian; Legavre, Thierry; Maximova, Siela N; Abrouk, Michael; Murat, Florent; Fouet, Olivier; Poulain, Julie; Ruiz, Manuel; Roguet, Yolande; Rodier-Goud, Maguy; Barbosa-Neto, Jose Fernandes; Sabot, Francois; Kudrna, Dave; Ammiraju, Jetty Siva S; Schuster, Stephan C; Carlson, John E; Sallet, Erika; Schiex, Thomas; Dievart, Anne; Kramer, Melissa; Gelley, Laura; Shi, Zi; Bérard, Aurélie; Viot, Christopher; Boccara, Michel; Risterucci, Ange Marie; Guignon, Valentin; Sabau, Xavier; Axtell, Michael J; Ma, Zhaorong; Zhang, Yufan; Brown, Spencer; Bourge, Mickael; Golser, Wolfgang; Song, Xiang; Clement, Didier; Rivallan, Ronan; Tahi, Mathias; Akaza, Joseph Moroh; Pitollat, Bertrand; Gramacho, Karina; D'Hont, Angélique; Brunel, Dominique; Infante, Diogenes; Kebe, Ismael; Costet, Pierre; Wing, Rod; McCombie, W Richard; Guiderdoni, Emmanuel; Quetier, Francis; Panaud, Olivier; Wincker, Patrick; Bocs, Stephanie; Lanaud, Claire

2011-02-01

We sequenced and assembled the draft genome of Theobroma cacao, an economically important tropical-fruit tree crop that is the source of chocolate. This assembly corresponds to 76% of the estimated genome size and contains almost all previously described genes, with 82% of these genes anchored on the 10 T. cacao chromosomes. Analysis of this sequence information highlighted specific expansion of some gene families during evolution, for example, flavonoid-related genes. It also provides a major source of candidate genes for T. cacao improvement. Based on the inferred paleohistory of the T. cacao genome, we propose an evolutionary scenario whereby the ten T. cacao chromosomes were shaped from an ancestor through eleven chromosome fusions.

High-resolution melt and morphological analyses of mealybugs (Hemiptera: Pseudococcidae) from cacao: tools for the control of Cacao swollen shoot virus spread.

PubMed

Wetten, Andy; Campbell, Colin; Allainguillaume, Joël

2016-03-01

Mealybugs (Hemiptera: Coccoidea: Pseudococcidae) are key vectors of badnaviruses, including Cacao swollen shoot virus (CSSV), the most damaging virus affecting cacao (Theobroma cacao L.). The effectiveness of mealybugs as virus vectors is species dependent, and it is therefore vital that CSSV resistance breeding programmes in cacao incorporate accurate mealybug identification. In this work, the efficacy of a CO1-based DNA barcoding approach to species identification was evaluated by screening a range of mealybugs collected from cacao in seven countries. Morphologically similar adult females were characterised by scanning electron microscopy, and then, following DNA extraction, were screened with CO1 barcoding markers. A high degree of CO1 sequence homology was observed for all 11 individual haplotypes, including those accessions from distinct geographical regions. This has allowed the design of a high-resolution melt (HRM) assay capable of rapid identification of the commonly encountered mealybug pests of cacao. HRM analysis readily differentiated between mealybug pests of cacao that cannot necessarily be identified by conventional morphological analysis. This new approach, therefore, has potential to facilitate breeding for resistance to CSSV and other mealybug-transmitted diseases. © 2015 Society of Chemical Industry.

In silico analysis of cacao (Theobroma cacao L.) genes that involved in pathogen and disease responses

NASA Astrophysics Data System (ADS)

Agung, Muhammad Budi; Budiarsa, I. Made; Suwastika, I. Nengah

2017-02-01

Cocoa bean is one of the main commodities from Indonesia for the world, which still have problem regarding yield degradation due to pathogens and disease attack. Developing robust cacao plant that genetically resistant to pathogen and disease attack is an ideal solution in over taking on this problem. The aim of this study was to identify Theobroma cacao genes on database of cacao genome that homolog to response genes of pathogen and disease attack in other plant, through in silico analysis. Basic information survey and gene identification were performed in GenBank and The Arabidopsis Information Resource database. The In silico analysis contains protein BLAST, homology test of each gene's protein candidates, and identification of homologue gene in Cacao Genome Database using data source "Theobroma cacao cv. Matina 1-6 v1.1" genome. Identification found that Thecc1EG011959t1 (EDS1), Thecc1EG006803t1 (EDS5), Thecc1EG013842t1 (ICS1), and Thecc1EG015614t1 (BG_PPAP) gene of Cacao Genome Database were Theobroma cacao genes that homolog to plant's resistance genes which highly possible to have similar functions of each gene's homologue gene.

Building genomic resources for Theobroma cacao

USDA-ARS?s Scientific Manuscript database

Theobroma cacao L (cacao: Malvaceae) is a small tree endemic to the Amazonian rain forest, where it most likely evolved. Cacao persists in populations of naturally outcrossing and inbreeding plants, as it is a species with a complex system of self-incompatibility, where only a fraction of the popul...

Soil microbial communities under cacao agroforestry and cover crop systems in Peru

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao) trees are grown in tropical regions worldwide for chocolate production. We studied the effects of agroforestry management systems and cover cropping on soil microbial communities under cacao in two different replicated field experiments in Peru. Two agroforestry systems, Imp...

A Larger Chocolate Chip-Development of a 15K Theobroma cacao L. SNP Array to Create High-Density Linkage Maps.

PubMed

Livingstone, Donald; Stack, Conrad; Mustiga, Guiliana M; Rodezno, Dayana C; Suarez, Carmen; Amores, Freddy; Feltus, Frank A; Mockaitis, Keithanne; Cornejo, Omar E; Motamayor, Juan C

2017-01-01

Cacao ( Theobroma cacao L.) is an important cash crop in tropical regions around the world and has a rich agronomic history in South America. As a key component in the cosmetic and confectionary industries, millions of people worldwide use products made from cacao, ranging from shampoo to chocolate. An Illumina Infinity II array was created using 13,530 SNPs identified within a small diversity panel of cacao. Of these SNPs, 12,643 derive from variation within annotated cacao genes. The genotypes of 3,072 trees were obtained, including two mapping populations from Ecuador. High-density linkage maps for these two populations were generated and compared to the cacao genome assembly. Phenotypic data from these populations were combined with the linkage maps to identify the QTLs for yield and disease resistance.

Trichoderma species form endophytic associations within Theobroma cacao trichomes.

PubMed

Bailey, Bryan A; Strem, Mary D; Wood, Delilah

2009-12-01

Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.

Unexpected Genome Variability at Multiple Loci Suggests Cacao Swollen Shoot Virus Comprises Multiple, Divergent Molecular Variants.

USDA-ARS?s Scientific Manuscript database

Cacao swollen shoot virus (CSSV) [Badnavirus, Caulimoviridae] causes swollen shoot disease of Theobroma cacao L. in West Africa. Since ~2000, various diagnostic tests have failed to detect CSSV in ~50-70% of symptomatic cacao plants, suggesting the possible emergence of new, previously uncharacteriz...

CACAO (THEOBROMA CACAO L.) GENETIC RESOURCES RESEARCH AT THE USDA-ARS TROPICAL AGRICULTURE RESEARCH STATION

USDA-ARS?s Scientific Manuscript database

The current USDA ARS Tropical Agriculture Research Station’s cacao (Theobroma cacao) collection consists of 154 clonally propagated accessions. Each accession is represented by six individual trees grafted on Amelonado rootstocks and planted in a completely randomized block design with three blocks...

Evaluation of soil amendments as a remediation alternative for cadmium contaminated soils under cacao plantations

USDA-ARS?s Scientific Manuscript database

Elevated plant-available cadmium (Cd) in soils results in contamination to cacao (Theobroma cacao L) beans. Effectiveness of vermicompost and zeolite in reducing available Cd in three cacao-growing soils was studied under laboratory conditions. Sorption-desorption experiments were conducted in soils...

Chemical and archaeological evidence for the earliest cacao beverages

PubMed Central

Henderson, John S.; Joyce, Rosemary A.; Hall, Gretchen R.; Hurst, W. Jeffrey; McGovern, Patrick E.

2007-01-01

Chemical analyses of residues extracted from pottery vessels from Puerto Escondido in what is now Honduras show that cacao beverages were being made there before 1000 B.C., extending the confirmed use of cacao back at least 500 years. The famous chocolate beverage served on special occasions in later times in Mesoamerica, especially by elites, was made from cacao seeds. The earliest cacao beverages consumed at Puerto Escondido were likely produced by fermenting the sweet pulp surrounding the seeds. PMID:18024588

Chemical and archaeological evidence for the earliest cacao beverages.

PubMed

Henderson, John S; Joyce, Rosemary A; Hall, Gretchen R; Hurst, W Jeffrey; McGovern, Patrick E

2007-11-27

Chemical analyses of residues extracted from pottery vessels from Puerto Escondido in what is now Honduras show that cacao beverages were being made there before 1000 B.C., extending the confirmed use of cacao back at least 500 years. The famous chocolate beverage served on special occasions in later times in Mesoamerica, especially by elites, was made from cacao seeds. The earliest cacao beverages consumed at Puerto Escondido were likely produced by fermenting the sweet pulp surrounding the seeds.

A Larger Chocolate Chip—Development of a 15K Theobroma cacao L. SNP Array to Create High-Density Linkage Maps

PubMed Central

Livingstone, Donald; Stack, Conrad; Mustiga, Guiliana M.; Rodezno, Dayana C.; Suarez, Carmen; Amores, Freddy; Feltus, Frank A.; Mockaitis, Keithanne; Cornejo, Omar E.; Motamayor, Juan C.

2017-01-01

Cacao (Theobroma cacao L.) is an important cash crop in tropical regions around the world and has a rich agronomic history in South America. As a key component in the cosmetic and confectionary industries, millions of people worldwide use products made from cacao, ranging from shampoo to chocolate. An Illumina Infinity II array was created using 13,530 SNPs identified within a small diversity panel of cacao. Of these SNPs, 12,643 derive from variation within annotated cacao genes. The genotypes of 3,072 trees were obtained, including two mapping populations from Ecuador. High-density linkage maps for these two populations were generated and compared to the cacao genome assembly. Phenotypic data from these populations were combined with the linkage maps to identify the QTLs for yield and disease resistance. PMID:29259608

Concentration of Cadmium in Cacao Beans and its Relationship with Soil Cadmium in Southern Ecuador

USDA-ARS?s Scientific Manuscript database

The concentration of cadmium (Cd) in cacao (Theobroma cacao, L.) beans above a critical level (0.6 mg kg-1 established by the European Union) has raised concerns of safety in the consumption of cacao-based chocolate (dark chocolate). Currently, little is available regarding Cd concentration in soil,...

A genetically anchored physical framework for Theobroma cacao cv. Matina 1-6

USDA-ARS?s Scientific Manuscript database

Background: Theobroma cacao (cacao tree) is the main ingredient in chocolate. World cocoa production is estimated to be 7 million tons in 2010 with an annual estimated average growth rate of 2.2%. This cacao bean production industry is currently under threat from a rise in fungal diseases includi...

Complex origin of Trinitario-type Theobroma cacao (Malvaceae) revealed using plastid genomics

USDA-ARS?s Scientific Manuscript database

Trinidad and Tobago has a long history of producing high quality cacao (Theobroma cacao L.) flavor, despite this industry having been threatened in the past by disease and changing economic fortunes. Cacao genotypes in Trinidad and Tobago are of a highly distinctive kind, the so-called “Trinitari...

Molecular characterization of previously elusive badnaviruses associated with symptomatic cacao in the New World.

USDA-ARS?s Scientific Manuscript database

ABSTRACT Cacao (Theobroma cacao L.) is a tropical tree cultivated for beans used to make chocolate. Distinct virus-like symptoms, referred to as the putative Cacao Trinidad virus (CTV) strains A and B, have been observed in Trinidad and Tobago since 1944, however, viral etiology had not been demon...

Making a chocolate chip: development and evaluation of a 6K SNP array for Theobroma cacao

PubMed Central

Livingstone, Donald; Royaert, Stefan; Stack, Conrad; Mockaitis, Keithanne; May, Greg; Farmer, Andrew; Saski, Christopher; Schnell, Ray; Kuhn, David; Motamayor, Juan Carlos

2015-01-01

Theobroma cacao, the key ingredient in chocolate production, is one of the world's most important tree fruit crops, with ∼4,000,000 metric tons produced across 50 countries. To move towards gene discovery and marker-assisted breeding in cacao, a single-nucleotide polymorphism (SNP) identification project was undertaken using RNAseq data from 16 diverse cacao cultivars. RNA sequences were aligned to the assembled transcriptome of the cultivar Matina 1-6, and 330,000 SNPs within coding regions were identified. From these SNPs, a subset of 6,000 high-quality SNPs were selected for inclusion on an Illumina Infinium SNP array: the Cacao6kSNP array. Using Cacao6KSNP array data from over 1,000 cacao samples, we demonstrate that our custom array produces a saturated genetic map and can be used to distinguish among even closely related genotypes. Our study enhances and expands the genetic resources available to the cacao research community, and provides the genome-scale set of tools that are critical for advancing breeding with molecular markers in an agricultural species with high genetic diversity. PMID:26070980

Gene structure, phylogeny and expression profile of the sucrose synthase gene family in cacao (Theobroma cacao L.).

PubMed

Li, Fupeng; Hao, Chaoyun; Yan, Lin; Wu, Baoduo; Qin, Xiaowei; Lai, Jianxiong; Song, Yinghui

2015-09-01

In higher plants, sucrose synthase (Sus, EC 2.4.1.13) is widely considered as a key enzyme involved in sucrose metabolism. Although, several paralogous genes encoding different isozymes of Sus have been identified and characterized in multiple plant genomes, to date detailed information about the Sus genes is lacking for cacao. This study reports the identification of six novel Sus genes from economically important cacao tree. Analyses of the gene structure and phylogeny of the Sus genes demonstrated evolutionary conservation in the Sus family across cacao and other plant species. The expression of cacao Sus genes was investigated via real-time PCR in various tissues, different developmental phases of leaf, flower bud and pod. The Sus genes exhibited distinct but partially redundant expression profiles in cacao, with TcSus1, TcSus5 and TcSus6, being the predominant genes in the bark with phloem, TcSus2 predominantly expressing in the seed during the stereotype stage. TcSus3 and TcSus4 were significantly detected more in the pod husk and seed coat along the pod development, and showed development dependent expression profiles in the cacao pod. These results provide new insights into the evolution, and basic information that will assist in elucidating the functions of cacao Sus gene family.

Cacao extracts suppress tryptophan degradation of mitogen-stimulated peripheral blood mononuclear cells.

PubMed

Jenny, M; Santer, E; Klein, A; Ledochowski, M; Schennach, H; Ueberall, F; Fuchs, D

2009-03-18

The fruits of Theobroma cacao L. (Sterculiaceae) have been used as food and a remedy for more than 4000 years. Today, about 100 therapeutic applications of cacao are described involving the gastrointestinal, nervous, cardiovascular and immune systems. Pro-inflammatory cytokine interferon-gamma and related biochemical pathways like tryptophan degradation by indoleamine 2,3-dioxygenase and neopterin formation are closely associated with the pathogenesis of such disorders. To determine the anti-inflammatory effect of cacao extracts on interferon-gamma and biochemical consequences in immunocompetent cells. Effects of aqueous or ethanolic extracts of cacao were examined on mitogen-induced human peripheral blood mononuclear cells (PBMC) of healthy donors and on lipopolysaccharide-stimulated myelomonocytic THP-1 cells. Antioxidant activity of extracts was determined by oxygen radical absorption capacity (ORAC) assay. In mitogen-stimulated PBMC, enhanced degradation of tryptophan, formation of neopterin and interferon-gamma were almost completely suppressed by the cacao extracts at doses of > or = 5 microg/mL. Cacao extracts had no effect on tryptophan degradation in lipopolysaccharide-stimulated THP-1 cells. There is a significant suppressive effect of cacao extracts on pro-inflammatory pathways in activated T-cells. Particularly the influence on indoleamine 2,3-dioxygenase could relate to some of the beneficial health effects ascribed to cacao.

Food of the gods: cure for humanity? A cultural history of the medicinal and ritual use of chocolate.

PubMed

Dillinger, T L; Barriga, P; Escárcega, S; Jimenez, M; Salazar Lowe, D; Grivetti, L E

2000-08-01

The medicinal use of cacao, or chocolate, both as a primary remedy and as a vehicle to deliver other medicines, originated in the New World and diffused to Europe in the mid 1500s. These practices originated among the Olmec, Maya and Mexica (Aztec). The word cacao is derived from Olmec and the subsequent Mayan languages (kakaw); the chocolate-related term cacahuatl is Nahuatl (Aztec language), derived from Olmec/Mayan etymology. Early colonial era documents included instructions for the medicinal use of cacao. The Badianus Codex (1552) noted the use of cacao flowers to treat fatigue, whereas the Florentine Codex (1590) offered a prescription of cacao beans, maize and the herb tlacoxochitl (Calliandra anomala) to alleviate fever and panting of breath and to treat the faint of heart. Subsequent 16th to early 20th century manuscripts produced in Europe and New Spain revealed >100 medicinal uses for cacao/chocolate. Three consistent roles can be identified: 1) to treat emaciated patients to gain weight; 2) to stimulate nervous systems of apathetic, exhausted or feeble patients; and 3) to improve digestion and elimination where cacao/chocolate countered the effects of stagnant or weak stomachs, stimulated kidneys and improved bowel function. Additional medical complaints treated with chocolate/cacao have included anemia, poor appetite, mental fatigue, poor breast milk production, consumption/tuberculosis, fever, gout, kidney stones, reduced longevity and poor sexual appetite/low virility. Chocolate paste was a medium used to administer drugs and to counter the taste of bitter pharmacological additives. In addition to cacao beans, preparations of cacao bark, oil (cacao butter), leaves and flowers have been used to treat burns, bowel dysfunction, cuts and skin irritations.

Colombia a Source of Cacao Genetic Diversity As Revealed by the Population Structure Analysis of Germplasm Bank of Theobroma cacao L.

PubMed Central

Osorio-Guarín, Jaime A.; Berdugo-Cely, Jhon; Coronado, Roberto Antonio; Zapata, Yeny Patricia; Quintero, Constanza; Gallego-Sánchez, Gerardo; Yockteng, Roxana

2017-01-01

Beans of the species Theobroma cacao L., also known as cacao, are the raw material to produce chocolate. Colombian cacao has been classified as a fine flavor cacao that represents the 5% of cacao world’s production. Colombian genetic resources from this species are conserved in ex situ and in-field germplasm banks, since T. cacao has recalcitrant seeds to desication and long-term storage. Currently, the collection of T. cacao of the Colombian Corporation of Agricultural Research (CORPOICA) has approximately 700 germplasm accessions. We conducted a molecular analysis of Corpoica’s cacao collection and a morphological characterization of some accessions with the goal to study its genetic diversity and population structure and, to select interesting accessions for the cacao’s breeding program. Phenotypic evaluation was performed based on 18 morphological traits and 4 biochemical traits. PCA analysis of morphological traits explained 60.6% of the total variation in seven components and 100% of the total variation of biochemical traits in four components, grouping the collection in 4 clusters for both variables. We explored 565 accessions from Corpoica’s germplasm and 252 accessions from reference populations using 96 single nucleotide polymorphism (SNP) molecular markers. Molecular patterns of cacao Corpoica’s collection were obtained amplifying specific alleles in a Fluidigm platform that used integrated circuits of fluids. Corpoica’s collection showed highest genetic diversity [Expected Heterozygosity (HE = 0.314), Observed Heterozygosity (HO = 0.353)] that is reduced when reference populations were included in the dataset (HE = 0.294, HO = 0.261). The collection was divided into four clusters based on population structure analysis. Cacao accessions from distinct groups showed some taxonomic concordance and reflected their geographic origins. For instance, accessions classified as Criollo were clearly differentiated in one group and we identified two new Colombian genetic groups. Using a number of allelic variations based on 87 SNP markers and 22 different morphological/biochemical traits, a core collection with a total of 232 accessions was selected as a primary genetic resource for cacao breeders. PMID:29209353

Impact Of Selfing On The Inference Of Demographic History From Whole Genomes In Theobroma cacao L.

USDA-ARS?s Scientific Manuscript database

Theobroma cacao L (cacao: Malvaceae) is a small tree found naturally in the Amazonian rain forest. An interesting feature of cacao is that it persists in populations of naturally outcrossing and inbreeding plants, as it is a species with a complex system of self-incompatibility, where a fraction of...

Optimization of a SNP assay for Genotyping Theobroma cacao under field conditions

USDA-ARS?s Scientific Manuscript database

The tropical tree crop Theobroma cacao L. is grown commercially for its beans, which are used in the production of cocoa butter and chocolate. Although the upper Amazon region is the center of origin for cacao, 70% of the world’s supply of cacao beans currently comes from small farms in West Africa...

Impact of soils and cropping systems on composition of mineral elements of dry cacao beans

USDA-ARS?s Scientific Manuscript database

In view of its high economic value, cacao (Theobroma cacao L.) researchers are seeking technological innovations that increase production and improve the quality of cacao beans. The objective of this study was to characterize the mineral (P, K, Ca, Mg, Si, Fe, Mn, Zn, Cu, Cd, Ba) composition of caca...

Single nucleotide polymorphisms from Theobroma cacao expressed sequence tags associated with witches' broom disease in cacao.

PubMed

Lima, L S; Gramacho, K P; Carels, N; Novais, R; Gaiotto, F A; Lopes, U V; Gesteira, A S; Zaidan, H A; Cascardo, J C M; Pires, J L; Micheli, F

2009-07-14

In order to increase the efficiency of cacao tree resistance to witches' broom disease, which is caused by Moniliophthora perniciosa (Tricholomataceae), we looked for molecular markers that could help in the selection of resistant cacao genotypes. Among the different markers useful for developing marker-assisted selection, single nucleotide polymorphisms (SNPs) constitute the most common type of sequence difference between alleles and can be easily detected by in silico analysis from expressed sequence tag libraries. We report the first detection and analysis of SNPs from cacao-M. perniciosa interaction expressed sequence tags, using bioinformatics. Selection based on analysis of these SNPs should be useful for developing cacao varieties resistant to this devastating disease.

Crude cacao Theobroma cacao extract reduces mutagenicity induced by benzo[a]pyrene through inhibition of CYP1A activity in vitro.

PubMed

Ohno, Marumi; Sakamoto, Kentaro Q; Ishizuka, Mayumi; Fujita, Shoichi

2009-08-01

Polyphenols have been shown to have potent antioxidant activity, and therefore, food containing polyphenols is expected to contribute to the prevention of cancer. However, food contains not only polyphenols but also various other constituents. We used the Ames test to investigate the effects of crude extracts of whole cacao products, which are known to be rich in polyphenols, on the mutagenicity of benzo[a]pyrene (B[a]P) in Salmonella typhimurium strain TA 98 and tert-butyl hydroperoxide (t-BuOOH) in S. typhimurium strain TA 102. B[a]P induces mutagenicity by metabolic activation and t-BuOOH induces it by generation of free radicals. While white chocolate did not modulate the numbers of revertant colonies produced by B[a]P treatment, milk chocolate and cacao powder extracts did. On the other hand, surprisingly, none of the cacao products tested affected the number of revertant colonies when t-BuOOH was used as the mutagen. At maximum concentration (13.25 mg cacao powder/ml), the crude cacao powder extract reduced ethoxyresorufin O-deethylase activity to 17.4% of the control, suggesting that whole cacao products inhibit cytochrome P450 (CYP) 1A activity. In conclusion, inhibition of CYP1A activity by cacao products may prevent DNA damage by reducing metabolic activation of carcinogens. Copyright 2009 John Wiley & Sons, Ltd.

QTL mapping for resistance to frosty pod and black pod diseases in an f1 population of Theobroma cacao L

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is a native crop of the Americas; however severe losses due to frosty pod (FP) [Moniliophthora roreri (Cif. and Par.)], and black pod (BP) [Phytophthora palmivora (Butl.) Butl.] have reduced cacao in the Americas to only 13.0% of world production. Agronomic practices to co...

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

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is native to the South American rainforest but it was domesticated in Mesoamerica. The relic Criollo cocoa in Belize has been well known in the premium chocolate market for its high-quality. Knowledge of genetic diversity in this variety is essential for efficient conserva...

The occurrence and frequency of Witches’ Broom Disease associated with Wild Cacao from the Upper Amazon of Peru

USDA-ARS?s Scientific Manuscript database

The Peruvian Amazon is within the center of origin and diversity for cacao (Theobroma cacao). One of the primary disease of cacao in Peru and Latin America is withes’ broom disease (WBD) caused by Moniliophthora perniciosa. The objective of this study was to determine the incidence of WBD in wild ca...

Making a chocolate chip: development and evaluation of a 6K SNP array for Theobroma cacao.

PubMed

Livingstone, Donald; Royaert, Stefan; Stack, Conrad; Mockaitis, Keithanne; May, Greg; Farmer, Andrew; Saski, Christopher; Schnell, Ray; Kuhn, David; Motamayor, Juan Carlos

2015-08-01

Theobroma cacao, the key ingredient in chocolate production, is one of the world's most important tree fruit crops, with ∼4,000,000 metric tons produced across 50 countries. To move towards gene discovery and marker-assisted breeding in cacao, a single-nucleotide polymorphism (SNP) identification project was undertaken using RNAseq data from 16 diverse cacao cultivars. RNA sequences were aligned to the assembled transcriptome of the cultivar Matina 1-6, and 330,000 SNPs within coding regions were identified. From these SNPs, a subset of 6,000 high-quality SNPs were selected for inclusion on an Illumina Infinium SNP array: the Cacao6kSNP array. Using Cacao6KSNP array data from over 1,000 cacao samples, we demonstrate that our custom array produces a saturated genetic map and can be used to distinguish among even closely related genotypes. Our study enhances and expands the genetic resources available to the cacao research community, and provides the genome-scale set of tools that are critical for advancing breeding with molecular markers in an agricultural species with high genetic diversity. © The Author 2015. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.

Accurate determination of genetic identity for a single cacao bean, using molecular markers with a nanofluidic system, ensures cocoa authentication.

PubMed

Fang, Wanping; Meinhardt, Lyndel W; Mischke, Sue; Bellato, Cláudia M; Motilal, Lambert; Zhang, Dapeng

2014-01-15

Cacao (Theobroma cacao L.), the source of cocoa, is an economically important tropical crop. One problem with the premium cacao market is contamination with off-types adulterating raw premium material. Accurate determination of the genetic identity of single cacao beans is essential for ensuring cocoa authentication. Using nanofluidic single nucleotide polymorphism (SNP) genotyping with 48 SNP markers, we generated SNP fingerprints for small quantities of DNA extracted from the seed coat of single cacao beans. On the basis of the SNP profiles, we identified an assumed adulterant variety, which was unambiguously distinguished from the authentic beans by multilocus matching. Assignment tests based on both Bayesian clustering analysis and allele frequency clearly separated all 30 authentic samples from the non-authentic samples. Distance-based principle coordinate analysis further supported these results. The nanofluidic SNP protocol, together with forensic statistical tools, is sufficiently robust to establish authentication and to verify gourmet cacao varieties. This method shows significant potential for practical application.

21 CFR 163.113 - Cocoa.

Code of Federal Regulations, 2011 CFR

2011-04-01

... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.113 Cocoa. (a... requirements for label declaration of ingredients for breakfast cocoa in § 163.112, except that the cacao fat...

40 CFR 180.628 - Chlorantraniliprole; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., subgroup 13-07B 2.5 Cacao bean 0.08 Cacao bean, chocolate 1.5 Cacao bean, cocoa powder 1.5 Cacao bean, roasted bean 0.8 Cactus 13 Canistel 4.0 Cattle, fat 0.5 Cattle, liver 0.3 Cattle, meat 0.1 Cattle, meat..., green bean 0.4 Coffee, instant 2.0 Corn, field, forage 14 Corn, field, grain 0.04 Corn, field, milled...

40 CFR 180.628 - Chlorantraniliprole; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., subgroup 13-07B 2.5 Cacao bean 0.08 Cacao bean, chocolate 1.5 Cacao bean, cocoa powder 1.5 Cacao bean, roasted bean 0.8 Cactus 13 Canistel 4.0 Cattle, fat 0.5 Cattle, meat 0.1 Cattle, meat byproducts 0.5 Cherimoya 4.0 Cherry, sweet 2.0 Cherry, tart 2.0 Citrus, dried pulp 14 Coffee, green bean 0.4 Coffee...

40 CFR 180.628 - Chlorantraniliprole; tolerances for residues.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., subgroup 13-07B 2.5 Cacao bean 0.08 Cacao bean, chocolate 1.5 Cacao bean, cocoa powder 1.5 Cacao bean, roasted bean 0.8 Cactus 13 Canistel 4.0 Cattle, fat 0.3 Cattle, liver 0.3 Cattle, meat 0.05 Cattle, meat..., green bean 0.4 Coffee, instant 2.0 Corn, field, forage 14 Corn, field, grain 0.04 Corn, field, milled...

Genome and secretome analysis of the hemibiotrophic fungal pathogen, Moniliophthora roreri, which causes frosty pod rot disease of cacao: mechanisms of the biotrophic and necrotrophic phases

USDA-ARS?s Scientific Manuscript database

Background: Moniliophthora roreri is the causal agent of Frosty pod rot (FPR) disease of Theobroma cacao, the source of chocolate and is one of the most destructive diseases of cacao in the Americas. This Basidiomycete only infects cacao pods and has an extended biotrophic phase lasting up to sixty ...

Dynamic changes in pod and fungal physiology associated with the shift from biotrophy to necrotrophy during the infection of Theobroma cacao by Moniliophthora roreri

USDA-ARS?s Scientific Manuscript database

Where it occurs in South and Central America, M. roreri (Mr) causes a destructive pod disease (frosty pod rot) on Theobroma cacao (cacao). Hand pollinated cacao pods were inoculated with Mr spores in the field and assessed for disease symptoms over a 90 day period. On average, pods showed symptoms o...

Phytophthora megakarya and P. palmivora, closely related causal agents of cacao black pod rot, underwent increases in genome sizes and gene numbers by different mechanisms

USDA-ARS?s Scientific Manuscript database

Phytophthora megakarya (Pmeg) and P. palmivora (Ppal) are closely related species causing black pod rot of cacao. While Ppal is a cosmopolitan plant pathogen, cacao is the only known host of importance for Pmeg. Pmeg is more virulent on cacao than Ppal. Therefore, we have sequenced both the Pmeg and...

Present spatial diversity patterns of Theobroma cacao L. in the neotropics reflect genetic differentiation in pleistocene refugia followed by human-influenced dispersal.

PubMed

Thomas, Evert; van Zonneveld, Maarten; Loo, Judy; Hodgkin, Toby; Galluzzi, Gea; van Etten, Jacob

2012-01-01

Cacao (Theobroma cacao L.) is indigenous to the Amazon basin, but is generally believed to have been domesticated in Mesoamerica for the production of chocolate beverage. However, cacao's distribution of genetic diversity in South America is also likely to reflect pre-Columbian human influences that were superimposed on natural processes of genetic differentiation. Here we present the results of a spatial analysis of the intra-specific diversity of cacao in Latin America, drawing on a dataset of 939 cacao trees genotypically characterized by means of 96 SSR markers. To assess continental diversity patterns we performed grid-based calculations of allelic richness, Shannon diversity and Nei gene diversity, and distinguished different spatially coherent genetic groups by means of cluster analysis. The highest levels of genetic diversity were observed in the Upper Amazon areas from southern Peru to the Ecuadorian Amazon and the border areas between Colombia, Peru and Brazil. On the assumption that the last glaciation (22,000-13,000 BP) had the greatest pre-human impact on the current distribution and diversity of cacao, we modeled the species' Pleistocene niche suitability and overlaid this with present-day diversity maps. The results suggest that cacao was already widely distributed in the Western Amazon before the onset of glaciation. During glaciations, cacao populations were likely to have been restricted to several refugia where they probably underwent genetic differentiation, resulting in a number of genetic clusters which are representative for, or closest related to, the original wild cacao populations. The analyses also suggested that genetic differentiation and geographical distribution of a number of other clusters seem to have been significantly affected by processes of human management and accompanying genetic bottlenecks. We discuss the implications of these results for future germplasm collection and in situ, on farm and ex situ conservation of cacao.

Cadmium bioaccumulation and gastric bioaccessibility in cacao: A field study in areas impacted by oil activities in Ecuador.

PubMed

Barraza, F; Schreck, E; Lévêque, T; Uzu, G; López, F; Ruales, J; Prunier, J; Marquet, A; Maurice, L

2017-10-01

Cacao from South America is especially used to produce premium quality chocolate. Although the European Food Safety Authority has not established a limit for cadmium (Cd) in chocolate raw material, recent studies demonstrate that Cd concentrations in cacao beans can reach levels higher than the legal limits for dark chocolate (0.8 mg kg -1 , effective January 1st, 2019). Despite the fact that the presence of Cd in agricultural soils is related to contamination by fertilizers, other potential sources must be considered in Ecuador. This field study was conducted to investigate Cd content in soils and cacao cultivated on Ecuadorian farms in areas impacted by oil activities. Soils, cacao leaves, and pod husks were collected from 31 farms in the northern Amazon and Pacific coastal regions exposed to oil production and refining and compared to two control areas. Human gastric bioaccessibility was determined in raw cacao beans and cacao liquor samples in order to assess potential health risks involved. Our results show that topsoils (0-20 cm) have higher Cd concentrations than deeper layers, exceeding the Ecuadorian legislation limit in 39% of the sampling sites. Cacao leaves accumulate more Cd than pod husks or beans but, nevertheless, 50% of the sampled beans have Cd contents above 0.8 mg kg -1 . Root-to-cacao transfer seems to be the main pathway of Cd uptake, which is not only regulated by physico-chemical soil properties but also agricultural practices. Additionally, natural Cd enrichment by volcanic inputs must not be neglected. Finally, Cd in cacao trees cannot be considered as a tracer of oil activities. Assuming that total Cd content and its bioaccessible fraction (up to 90%) in cacao beans and liquor is directly linked to those in chocolate, the health risk associated with Cd exposure varies from low to moderate. Copyright © 2017 Elsevier Ltd. All rights reserved.

TcCYPR04, a Cacao Papain-Like Cysteine-Protease Detected in Senescent and Necrotic Tissues Interacts with a Cystatin TcCYS4

PubMed Central

Cardoso, Thyago Hermylly Santana; Freitas, Ana Camila Oliveira; Andrade, Bruno Silva; de Sousa, Aurizangela Oliveira; Santiago, André da Silva; Koop, Daniela Martins; Gramacho, Karina Peres; Alvim, Fátima Cerqueira; Micheli, Fabienne; Pirovani, Carlos Priminho

2015-01-01

The interaction amongst papain-like cysteine-proteases (PLCP) and their substrates and inhibitors, such as cystatins, can be perceived as part of the molecular battlefield in plant-pathogen interaction. In cacao, four cystatins were identified and characterized by our group. We identified 448 proteases in cacao genome, whereof 134 were cysteine-proteases. We expressed in Escherichia coli a PLCP from cacao, named TcCYSPR04. Immunoblottings with anti-TcCYSPR04 exhibited protein increases during leaf development. Additional isoforms of TcCYSPR04 appeared in senescent leaves and cacao tissues infected by Moniliophthora perniciosa during the transition from the biotrophic to the saprophytic phase. TcCYSPR04 was induced in the apoplastic fluid of Catongo and TSH1188 cacao genotypes, susceptible and resistant to M. perniciosa, respectively, but greater intensity and additional isoforms were observed in TSH1188. The fungal protein MpNEP induced PLCP isoform expression in tobacco leaves, according to the cross reaction with anti-TcCYSPR04. Several protein isoforms were detected at 72 hours after treatment with MpNEP. We captured an active PLCP from cacao tissues, using a recombinant cacao cystatin immobilized in CNBr-Sepharose. Mass spectrometry showed that this protein corresponds to TcCYSPR04. A homology modeling was obtained for both proteins. In order to become active, TcCYSPR04 needs to lose its inhibitory domain. Molecular docking showed the physical-chemical complementarities of the interaction between the cacao enzyme and its inhibitor. We propose that TcCYSPR04 and its interactions with cacao cystatins are involved in the senescence and necrosis events related to witches’ broom symptoms. This molecular interaction may be the target for future interventions to control witches' broom disease. PMID:26641247

TcCYPR04, a Cacao Papain-Like Cysteine-Protease Detected in Senescent and Necrotic Tissues Interacts with a Cystatin TcCYS4.

PubMed

Cardoso, Thyago Hermylly Santana; Freitas, Ana Camila Oliveira; Andrade, Bruno Silva; Sousa, Aurizangela Oliveira de; Santiago, André da Silva; Koop, Daniela Martins; Gramacho, Karina Peres; Alvim, Fátima Cerqueira; Micheli, Fabienne; Pirovani, Carlos Priminho

2015-01-01

The interaction amongst papain-like cysteine-proteases (PLCP) and their substrates and inhibitors, such as cystatins, can be perceived as part of the molecular battlefield in plant-pathogen interaction. In cacao, four cystatins were identified and characterized by our group. We identified 448 proteases in cacao genome, whereof 134 were cysteine-proteases. We expressed in Escherichia coli a PLCP from cacao, named TcCYSPR04. Immunoblottings with anti-TcCYSPR04 exhibited protein increases during leaf development. Additional isoforms of TcCYSPR04 appeared in senescent leaves and cacao tissues infected by Moniliophthora perniciosa during the transition from the biotrophic to the saprophytic phase. TcCYSPR04 was induced in the apoplastic fluid of Catongo and TSH1188 cacao genotypes, susceptible and resistant to M. perniciosa, respectively, but greater intensity and additional isoforms were observed in TSH1188. The fungal protein MpNEP induced PLCP isoform expression in tobacco leaves, according to the cross reaction with anti-TcCYSPR04. Several protein isoforms were detected at 72 hours after treatment with MpNEP. We captured an active PLCP from cacao tissues, using a recombinant cacao cystatin immobilized in CNBr-Sepharose. Mass spectrometry showed that this protein corresponds to TcCYSPR04. A homology modeling was obtained for both proteins. In order to become active, TcCYSPR04 needs to lose its inhibitory domain. Molecular docking showed the physical-chemical complementarities of the interaction between the cacao enzyme and its inhibitor. We propose that TcCYSPR04 and its interactions with cacao cystatins are involved in the senescence and necrosis events related to witches' broom symptoms. This molecular interaction may be the target for future interventions to control witches' broom disease.

Theobroma cacao L. pathogenesis-related gene tandem array members show diverse expression dynamics in response to pathogen colonization.

PubMed

Fister, Andrew S; Mejia, Luis C; Zhang, Yufan; Herre, Edward Allen; Maximova, Siela N; Guiltinan, Mark J

2016-05-17

The pathogenesis-related (PR) group of proteins are operationally defined as polypeptides that increase in concentration in plant tissues upon contact with a pathogen. To date, 17 classes of highly divergent proteins have been described that act through multiple mechanisms of pathogen resistance. Characterizing these families in cacao, an economically important tree crop, and comparing the families to those in other species, is an important step in understanding cacao's immune response. Using publically available resources, all members of the 17 recognized pathogenesis-related gene families in the genome of Theobroma cacao were identified and annotated resulting in a set of ~350 members in both published cacao genomes. Approximately 50 % of these genes are organized in tandem arrays scattered throughout the genome. This feature was observed in five additional plant taxa (three dicots and two monocots), suggesting that tandem duplication has played an important role in the evolution of the PR genes in higher plants. Expression profiling captured the dynamics and complexity of PR genes expression at basal levels and after induction by two cacao pathogens (the oomycete, Phytophthora palmivora, and the fungus, Colletotrichum theobromicola), identifying specific genes within families that are more responsive to pathogen challenge. Subsequent qRT-PCR validated the induction of several PR-1, PR-3, PR-4, and PR-10 family members, with greater than 1000 fold induction detected for specific genes. We describe candidate genes that are likely to be involved in cacao's defense against Phytophthora and Colletotrichum infection and could be potentially useful for marker-assisted selection for breeding of disease resistant cacao varieties. The data presented here, along with existing cacao-omics resources, will enable targeted functional genetic screening of defense genes likely to play critical functions in cacao's defense against its pathogens.

Shade tree diversity and aboveground carbon stocks in Theobroma cacao agroforestry systems: implications for REDD+ implementation in a West African cacao landscape.

PubMed

Dawoe, Evans; Asante, Winston; Acheampong, Emmanuel; Bosu, Paul

2016-12-01

The promotion of cacao agroforestry is one of the ways of diversifying farmer income and creating incentives through their inclusion in REDD+ interventions. We estimated the aboveground carbon stocks in cacao and shade trees, determined the floristic diversity of shade trees and explored the possibility of implementing REDD+ interventions in cacao landscapes. Using replicated multi-site transect approach, data were collected from nine 1-ha plots established on 5 km long transects in ten cacao growing districts in Ghana West Africa. Biomass of cacao and shade trees was determined using allometric equations. One thousand four hundred and one (1401) shade trees comprising 109 species from 33 families were recorded. Total number of species ranged from 34 to 49. Newbouldia laevis (Bignoniacea) was the most frequently occurring specie and constituted 43.2 % of all shade trees. The most predominant families were Sterculiaceae and Moraceae (10 species each), followed by Meliaceae and Mimosaceae (8 species each) and Caesalpiniacaea (6 species). Shannon diversity indices (H', H max and J') and species richness were low compared to other similar studies. Shade tree densities ranged from 16.2 ± 3.0 to 22.8 ± 1.7 stems ha -1 and differed significantly between sites. Carbon stocks of shade trees differed between sites but were similar in cacao trees. The average C stock in cacao trees was 7.45 ± 0.41 Mg C ha -1 compared with 8.32 ± 1.15 Mg C ha -1 in the shade trees. Cacao landscapes in Ghana have the potential of contributing to forest carbon stocks enhancement by increasing the stocking density of shade trees to recommended levels.

21 CFR 163.114 - Lowfat cocoa.

Code of Federal Regulations, 2011 CFR

2011-04-01

... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.114 Lowfat cocoa. (a... that the cacao fat content is less than 10 percent by weight, as determined by the method prescribed in...

Diversity of endophytic fungal community of cacao (Theobroma cacao L.) and biological control of Crinipellis perniciosa, causal agent of Witches' Broom Disease

PubMed Central

2005-01-01

The basidiomycete fungus Crinipellis perniciosa (Stahel) Singer is the causal agent of Witches' Broom Disease of Cacao (Theobroma cacao L.) which is the main factor limiting cacao production in the Americas. Pod losses of up to 90% are experienced in affected areas as evidenced by the 50% drop in production in Bahia province, Brazil following the arrival of the C. perniciosa in the area in 1989. The disease has proven particularly difficult to control and many farmers in affected areas have given up cacao cultivation. In order to evaluate the potential of endophytes as a biological control agent of this phytopathogen, the endophytic fungal community of resistant and susceptible cacao plants as well as affected branches was studied between 2001 and 2002. The fungal community was identified by morphological traits and rDNA sequencing as belonging to the genera Acremonium, Blastomyces, Botryosphaeria, Cladosporium, Colletotrichum, Cordyceps, Diaporthe, Fusarium, Geotrichum, Gibberella, Gliocladium, Lasiodiplodia, Monilochoetes, Nectria, Pestalotiopsis, Phomopsis, Pleurotus, Pseudofusarium, Rhizopycnis, Syncephalastrum, Trichoderma, Verticillium and Xylaria. These fungi were evaluated both in vitro and in vivo by their ability to inhibit C. perniciosa. Among these, some were identified as potential antagonists, but only one fungus (Gliocladium catenulatum) reduced the incidence of Witches' Broom Disease in cacao seedlings to 70%. PMID:15951847

Discovering terroir in the world of chocolate.

PubMed

Nesto, Bill

2010-01-01

The author investigates the applicability of the word “terroir” to chocolate. As a Master of Wine, wine journalist, and wine educator, the author has tried to understand how “terroir,” the environmental and human factors associated with growing vines and making wine, impacts the flavor of wine. Comparing and contrasting viticulture and winemaking to cacao farming and chocolate manufacture, the author analyzes to what degree terroir could be a concept that informs chocolate appreciation. He notes that the great distances between cacao farms and factories encourage the perception of cacao and chocolate as commodities. He observes that the varietal and origin nomenclature of cacao can be at worst misleading and generally lacks clarity and precision. He shows how the many steps that transform cacao into chocolate threaten the expression of terroir in the final product. Yet he acknowledges that there could be a basis for use of the word in the world of cacao and chocolate.

C and N Content in Density Fractions of Whole Soil and Soil Size Fraction Under Cacao Agroforestry Systems and Natural Forest in Bahia, Brazil

NASA Astrophysics Data System (ADS)

Rita, Joice Cleide O.; Gama-Rodrigues, Emanuela Forestieri; Gama-Rodrigues, Antonio Carlos; Polidoro, Jose Carlos; Machado, Regina Cele R.; Baligar, Virupax C.

2011-07-01

Agroforestry systems (AFSs) have an important role in capturing above and below ground soil carbon and play a dominant role in mitigation of atmospheric CO2. Attempts has been made here to identify soil organic matter fractions in the cacao-AFSs that have different susceptibility to microbial decomposition and further represent the basis of understanding soil C dynamics. The objective of this study was to characterize the organic matter density fractions and soil size fractions in soils of two types of cacao agroforestry systems and to compare with an adjacent natural forest in Bahia, Brazil. The land-use systems studied were: (1) a 30-year-old stand of natural forest with cacao (cacao cabruca), (2) a 30-year-old stand of cacao with Erythrina glauca as shade trees (cacao + erythrina), and (3) an adjacent natural forest without cacao. Soil samples were collected from 0-10 cm depth layer in reddish-yellow Oxisols. Soil samples was separated by wet sieving into five fraction-size classes (>2000 μm, 1000-2000 μm, 250-1000 μm, 53-250 μm, and <53 μm). C and N accumulated in to the light (free- and intra-aggregate density fractions) and heavy fractions of whole soil and soil size fraction were determined. Soil size fraction obtained in cacao AFS soils consisted mainly (65 %) of mega-aggregates (>2000 μm) mixed with macroaggregates (32-34%), and microaggregates (1-1.3%). Soil organic carbon (SOC) and total N content increased with increasing soil size fraction in all land-use systems. Organic C-to-total N ratio was higher in the macroaggregate than in the microaggregate. In general, in natural forest and cacao cabruca the contribution of C and N in the light and heavy fractions was similar. However, in cacao + erythrina the heavy fraction was the most common and contributed 67% of C and 63% of N. Finding of this study shows that the majority of C and N in all three systems studied are found in macroaggregates, particularly in the 250-1000 μm size aggregate class. The heavy fraction was the most common organic matter fraction in these soils. Thus, in mature cacao AFS on highly weathered soils the main mechanisms of C stabilization could be the physical protection within macroaggregate structures thereby minimizing the impact of conversion of forest to cacao AFS.

C and N content in density fractions of whole soil and soil size fraction under cacao agroforestry systems and natural forest in Bahia, Brazil.

PubMed

Rita, Joice Cleide O; Gama-Rodrigues, Emanuela Forestieri; Gama-Rodrigues, Antonio Carlos; Polidoro, Jose Carlos; Machado, Regina Cele R; Baligar, Virupax C

2011-07-01

Agroforestry systems (AFSs) have an important role in capturing above and below ground soil carbon and play a dominant role in mitigation of atmospheric CO(2). Attempts has been made here to identify soil organic matter fractions in the cacao-AFSs that have different susceptibility to microbial decomposition and further represent the basis of understanding soil C dynamics. The objective of this study was to characterize the organic matter density fractions and soil size fractions in soils of two types of cacao agroforestry systems and to compare with an adjacent natural forest in Bahia, Brazil. The land-use systems studied were: (1) a 30-year-old stand of natural forest with cacao (cacao cabruca), (2) a 30-year-old stand of cacao with Erythrina glauca as shade trees (cacao + erythrina), and (3) an adjacent natural forest without cacao. Soil samples were collected from 0-10 cm depth layer in reddish-yellow Oxisols. Soil samples was separated by wet sieving into five fraction-size classes (>2000 μm, 1000-2000 μm, 250-1000 μm, 53-250 μm, and <53 μm). C and N accumulated in to the light (free- and intra-aggregate density fractions) and heavy fractions of whole soil and soil size fraction were determined. Soil size fraction obtained in cacao AFS soils consisted mainly (65 %) of mega-aggregates (>2000 μm) mixed with macroaggregates (32-34%), and microaggregates (1-1.3%). Soil organic carbon (SOC) and total N content increased with increasing soil size fraction in all land-use systems. Organic C-to-total N ratio was higher in the macroaggregate than in the microaggregate. In general, in natural forest and cacao cabruca the contribution of C and N in the light and heavy fractions was similar. However, in cacao + erythrina the heavy fraction was the most common and contributed 67% of C and 63% of N. Finding of this study shows that the majority of C and N in all three systems studied are found in macroaggregates, particularly in the 250-1000 μm size aggregate class. The heavy fraction was the most common organic matter fraction in these soils. Thus, in mature cacao AFS on highly weathered soils the main mechanisms of C stabilization could be the physical protection within macroaggregate structures thereby minimizing the impact of conversion of forest to cacao AFS.

Differential expression of jasmonate biosynthesis genes in cacao genotypes contrasting for resistance against Moniliophthora perniciosa.

PubMed

Litholdo, Celso G; Leal, Gildemberg A; Albuquerque, Paulo S B; Figueira, Antonio

2015-10-01

The resistance mechanism of cacao against M. perniciosa is likely to be mediated by JA/ET-signaling pathways due to the preferential TcAOS and TcSAM induction in a resistant genotype. The basidiomycete Moniliophthora perniciosa causes a serious disease in cacao (Theobroma cacao L.), and the use of resistant varieties is the only sustainable long-term solution. Cacao resistance against M. perniciosa is characterized by pathogen growth inhibition with reduced colonization and an attenuation of disease symptoms, suggesting a regulation by jasmonate (JA)/ethylene (ET) signaling pathways. The hypothesis that genes involved in JA biosynthesis would be active in the interaction of T. cacao and M. perniciosa was tested here. The cacao JA-related genes were evaluated for their relative quantitative expression in susceptible and resistant genotypes upon the exogenous application of ET, methyl-jasmonate (MJ), and salicylic acid (SA), or after M. perniciosa inoculation. MJ treatment triggered changes in the expression of genes involved in JA biosynthesis, indicating that the mechanism of positive regulation by exogenous MJ application occurs in cacao. However, a higher induction of these genes was observed in the susceptible genotype. Further, a contrast in JA-related transcriptional expression was detected between susceptible and resistant plants under M. perniciosa infection, with the induction of the allene oxide synthase gene (TcAOS), which encodes a key enzyme in the JA biosynthesis pathway in the resistant genotype. Altogether, this work provides additional evidences that the JA-dependent signaling pathway is modulating the defense response against M. perniciosa in a cacao-resistant genotype.

Cocoa/Cotton Comparative Genomics

USDA-ARS?s Scientific Manuscript database

With genome sequence from two members of the Malvaceae family recently made available, we are exploring syntenic relationships, gene content, and evolutionary trajectories between the cacao and cotton genomes. An assembly of cacao (Theobroma cacao) using Illumina and 454 sequence technology yielded ...

Cacao use and the San Lorenzo Olmec.

PubMed

Powis, Terry G; Cyphers, Ann; Gaikwad, Nilesh W; Grivetti, Louis; Cheong, Kong

2011-05-24

Mesoamerican peoples had a long history of cacao use--spanning more than 34 centuries--as confirmed by previous identification of cacao residues on archaeological pottery from Paso de la Amada on the Pacific Coast and the Olmec site of El Manatí on the Gulf Coast. Until now, comparable evidence from San Lorenzo, the premier Olmec capital, was lacking. The present study of theobromine residues confirms the continuous presence and use of cacao products at San Lorenzo between 1800 and 1000 BCE, and documents assorted vessels forms used in its preparation and consumption. One elite context reveals cacao use as part of a mortuary ritual for sacrificial victims, an event that occurred during the height of San Lorenzo's power.

Towards the understanding of the cocoa transcriptome: Production and analysis of an exhaustive dataset of ESTs of Theobroma cacao L. generated from various tissues and under various conditions

PubMed Central

Argout, Xavier; Fouet, Olivier; Wincker, Patrick; Gramacho, Karina; Legavre, Thierry; Sabau, Xavier; Risterucci, Ange Marie; Da Silva, Corinne; Cascardo, Julio; Allegre, Mathilde; Kuhn, David; Verica, Joseph; Courtois, Brigitte; Loor, Gaston; Babin, Regis; Sounigo, Olivier; Ducamp, Michel; Guiltinan, Mark J; Ruiz, Manuel; Alemanno, Laurence; Machado, Regina; Phillips, Wilberth; Schnell, Ray; Gilmour, Martin; Rosenquist, Eric; Butler, David; Maximova, Siela; Lanaud, Claire

2008-01-01

Background Theobroma cacao L., is a tree originated from the tropical rainforest of South America. It is one of the major cash crops for many tropical countries. T. cacao is mainly produced on smallholdings, providing resources for 14 million farmers. Disease resistance and T. cacao quality improvement are two important challenges for all actors of cocoa and chocolate production. T. cacao is seriously affected by pests and fungal diseases, responsible for more than 40% yield losses and quality improvement, nutritional and organoleptic, is also important for consumers. An international collaboration was formed to develop an EST genomic resource database for cacao. Results Fifty-six cDNA libraries were constructed from different organs, different genotypes and different environmental conditions. A total of 149,650 valid EST sequences were generated corresponding to 48,594 unigenes, 12,692 contigs and 35,902 singletons. A total of 29,849 unigenes shared significant homology with public sequences from other species. Gene Ontology (GO) annotation was applied to distribute the ESTs among the main GO categories. A specific information system (ESTtik) was constructed to process, store and manage this EST collection allowing the user to query a database. To check the representativeness of our EST collection, we looked for the genes known to be involved in two different metabolic pathways extensively studied in other plant species and important for T. cacao qualities: the flavonoid and the terpene pathways. Most of the enzymes described in other crops for these two metabolic pathways were found in our EST collection. A large collection of new genetic markers was provided by this ESTs collection. Conclusion This EST collection displays a good representation of the T. cacao transcriptome, suitable for analysis of biochemical pathways based on oligonucleotide microarrays derived from these ESTs. It will provide numerous genetic markers that will allow the construction of a high density gene map of T. cacao. This EST collection represents a unique and important molecular resource for T. cacao study and improvement, facilitating the discovery of candidate genes for important T. cacao trait variation. PMID:18973681

Use of Trichoderma fungi in spray solutions to reduce Moniliophthora roreri infection of Theobroma cacao fruits in Northeastern Costa Rica.

PubMed

Seng, John; Herrera, Geovanny; Vaughan, Christopher S; McCoy, Michael B

2014-09-01

Cacao (Theobroma cacao) is an important cash crop in tropical climates such as that of Latin America. Over the past several decades, the infection of cultivated cacao by Moniliophthllora roreri, known commonly as "monilia", has significantly hindered cacao production in Latin America. Studies have proposed the use of Trichoderma sp. fungi in biocontrol treatments to prevent and reduce monilia infection, yet tests of Trichoderma-containing spray treatments on cacao agroforests have produced mixed results. Researchers and agricultural workers have suggested that addition of soil, fly ash, or other carbon sources to a Trichoderma spray may improve its efficacy in fighting monilia. To test these suggestions, we designed a series of spray mixtures including Thichoderma cultures, soil, and all necessary controls. We applied the spray mixtures to 80 cacao trees (20 trees for each of four resistant-selected clones to monilia) at the FINMAC organic cacao plantation in Pueblo Nuevo de Guacimo, Limón Province, in northeastern Costa Rica in March-April 2013. Five treatments were applied (control, water, water plus sterilized soil, water plus Trichoderma, and water plus sterilized soil plus Trichoderma). Each treatment was applied to four trees of each clone. We monitored the incidence of monilia infection under each spray treatment over the course of 35d. We found that spraying entire cacao trees two times with a mixture containing Trichoderma and sterilized soil significantly reduced the incidence of monilia infection by 11% (p ≤ 0.05) in only 35d, as compared to the control. This reduction in loss of cacao pods translates into an increase of plantation mean productivity of 1,500 kg dried beans/ha by 198 kg/ha up to 1,698 kg/ha or by a total increase over the whole 110 ha plantation by 21,780 kg. We propose that using such an antifungal spray over the whole course of a crop cycle (120 days) would decrease infection incidence even more. Application of this fungal control measure has the potential of revitalizing the production of cacao in the region.

Towards the understanding of the cocoa transcriptome: Production and analysis of an exhaustive dataset of ESTs of Theobroma cacao L. generated from various tissues and under various conditions.

PubMed

Argout, Xavier; Fouet, Olivier; Wincker, Patrick; Gramacho, Karina; Legavre, Thierry; Sabau, Xavier; Risterucci, Ange Marie; Da Silva, Corinne; Cascardo, Julio; Allegre, Mathilde; Kuhn, David; Verica, Joseph; Courtois, Brigitte; Loor, Gaston; Babin, Regis; Sounigo, Olivier; Ducamp, Michel; Guiltinan, Mark J; Ruiz, Manuel; Alemanno, Laurence; Machado, Regina; Phillips, Wilberth; Schnell, Ray; Gilmour, Martin; Rosenquist, Eric; Butler, David; Maximova, Siela; Lanaud, Claire

2008-10-30

Theobroma cacao L., is a tree originated from the tropical rainforest of South America. It is one of the major cash crops for many tropical countries. T. cacao is mainly produced on smallholdings, providing resources for 14 million farmers. Disease resistance and T. cacao quality improvement are two important challenges for all actors of cocoa and chocolate production. T. cacao is seriously affected by pests and fungal diseases, responsible for more than 40% yield losses and quality improvement, nutritional and organoleptic, is also important for consumers. An international collaboration was formed to develop an EST genomic resource database for cacao. Fifty-six cDNA libraries were constructed from different organs, different genotypes and different environmental conditions. A total of 149,650 valid EST sequences were generated corresponding to 48,594 unigenes, 12,692 contigs and 35,902 singletons. A total of 29,849 unigenes shared significant homology with public sequences from other species.Gene Ontology (GO) annotation was applied to distribute the ESTs among the main GO categories.A specific information system (ESTtik) was constructed to process, store and manage this EST collection allowing the user to query a database.To check the representativeness of our EST collection, we looked for the genes known to be involved in two different metabolic pathways extensively studied in other plant species and important for T. cacao qualities: the flavonoid and the terpene pathways. Most of the enzymes described in other crops for these two metabolic pathways were found in our EST collection.A large collection of new genetic markers was provided by this ESTs collection. This EST collection displays a good representation of the T. cacao transcriptome, suitable for analysis of biochemical pathways based on oligonucleotide microarrays derived from these ESTs. It will provide numerous genetic markers that will allow the construction of a high density gene map of T. cacao. This EST collection represents a unique and important molecular resource for T. cacao study and improvement, facilitating the discovery of candidate genes for important T. cacao trait variation.

Present Spatial Diversity Patterns of Theobroma cacao L. in the Neotropics Reflect Genetic Differentiation in Pleistocene Refugia Followed by Human-Influenced Dispersal

PubMed Central

Thomas, Evert; van Zonneveld, Maarten; Loo, Judy; Hodgkin, Toby; Galluzzi, Gea; van Etten, Jacob

2012-01-01

Cacao (Theobroma cacao L.) is indigenous to the Amazon basin, but is generally believed to have been domesticated in Mesoamerica for the production of chocolate beverage. However, cacao’s distribution of genetic diversity in South America is also likely to reflect pre-Columbian human influences that were superimposed on natural processes of genetic differentiation. Here we present the results of a spatial analysis of the intra-specific diversity of cacao in Latin America, drawing on a dataset of 939 cacao trees genotypically characterized by means of 96 SSR markers. To assess continental diversity patterns we performed grid-based calculations of allelic richness, Shannon diversity and Nei gene diversity, and distinguished different spatially coherent genetic groups by means of cluster analysis. The highest levels of genetic diversity were observed in the Upper Amazon areas from southern Peru to the Ecuadorian Amazon and the border areas between Colombia, Peru and Brazil. On the assumption that the last glaciation (22,000–13,000 BP) had the greatest pre-human impact on the current distribution and diversity of cacao, we modeled the species’ Pleistocene niche suitability and overlaid this with present-day diversity maps. The results suggest that cacao was already widely distributed in the Western Amazon before the onset of glaciation. During glaciations, cacao populations were likely to have been restricted to several refugia where they probably underwent genetic differentiation, resulting in a number of genetic clusters which are representative for, or closest related to, the original wild cacao populations. The analyses also suggested that genetic differentiation and geographical distribution of a number of other clusters seem to have been significantly affected by processes of human management and accompanying genetic bottlenecks. We discuss the implications of these results for future germplasm collection and in situ, on farm and ex situ conservation of cacao. PMID:23112832

Production and Robustness of a Cacao Agroecosystem: Effects of Two Contrasting Types of Management Strategies

PubMed Central

Sabatier, Rodolphe; Wiegand, Kerstin; Meyer, Katrin

2013-01-01

Ecological intensification, i.e. relying on ecological processes to replace chemical inputs, is often presented as the ideal alternative to conventional farming based on an intensive use of chemicals. It is said to both maintain high yield and provide more robustness to the agroecosystem. However few studies compared the two types of management with respect to their consequences for production and robustness toward perturbation. In this study our aim is to assess productive performance and robustness toward diverse perturbations of a Cacao agroecosystem managed with two contrasting groups of strategies: one group of strategies relying on a high level of pesticides and a second relying on low levels of pesticides. We conducted this study using a dynamical model of a Cacao agroecosystem that includes Cacao production dynamics, and dynamics of three insects: a pest (the Cacao Pod Borer, Conopomorpha cramerella) and two characteristic but unspecified beneficial insects (a pollinator of Cacao and a parasitoid of the Cacao Pod Borer). Our results showed two opposite behaviors of the Cacao agroecosystem depending on its management, i.e. an agroecosystem relying on a high input of pesticides and showing low ecosystem functioning and an agroecosystem with low inputs, relying on a high functioning of the ecosystem. From the production point of view, no type of management clearly outclassed the other and their ranking depended on the type of pesticide used. From the robustness point of view, the two types of managements performed differently when subjected to different types of perturbations. Ecologically intensive systems were more robust to pest outbreaks and perturbations related to pesticide characteristics while chemically intensive systems were more robust to Cacao production and management-related perturbation. PMID:24312469

Production and robustness of a Cacao agroecosystem: effects of two contrasting types of management strategies.

PubMed

Sabatier, Rodolphe; Wiegand, Kerstin; Meyer, Katrin

2013-01-01

Ecological intensification, i.e. relying on ecological processes to replace chemical inputs, is often presented as the ideal alternative to conventional farming based on an intensive use of chemicals. It is said to both maintain high yield and provide more robustness to the agroecosystem. However few studies compared the two types of management with respect to their consequences for production and robustness toward perturbation. In this study our aim is to assess productive performance and robustness toward diverse perturbations of a Cacao agroecosystem managed with two contrasting groups of strategies: one group of strategies relying on a high level of pesticides and a second relying on low levels of pesticides. We conducted this study using a dynamical model of a Cacao agroecosystem that includes Cacao production dynamics, and dynamics of three insects: a pest (the Cacao Pod Borer, Conopomorpha cramerella) and two characteristic but unspecified beneficial insects (a pollinator of Cacao and a parasitoid of the Cacao Pod Borer). Our results showed two opposite behaviors of the Cacao agroecosystem depending on its management, i.e. an agroecosystem relying on a high input of pesticides and showing low ecosystem functioning and an agroecosystem with low inputs, relying on a high functioning of the ecosystem. From the production point of view, no type of management clearly outclassed the other and their ranking depended on the type of pesticide used. From the robustness point of view, the two types of managements performed differently when subjected to different types of perturbations. Ecologically intensive systems were more robust to pest outbreaks and perturbations related to pesticide characteristics while chemically intensive systems were more robust to Cacao production and management-related perturbation.

Transient Expression of CRISPR/Cas9 Machinery Targeting TcNPR3 Enhances Defense Response in Theobroma cacao.

PubMed

Fister, Andrew S; Landherr, Lena; Maximova, Siela N; Guiltinan, Mark J

2018-01-01

Theobroma cacao , the source of cocoa, suffers significant losses to a variety of pathogens resulting in reduced incomes for millions of farmers in developing countries. Development of disease resistant cacao varieties is an essential strategy to combat this threat, but is limited by sources of genetic resistance and the slow generation time of this tropical tree crop. In this study, we present the first application of genome editing technology in cacao, using Agrobacterium-mediated transient transformation to introduce CRISPR/Cas9 components into cacao leaves and cotyledon cells. As a first proof of concept, we targeted the cacao Non-Expressor of Pathogenesis-Related 3 (TcNPR3) gene, a suppressor of the defense response. After demonstrating activity of designed single-guide RNAs (sgRNA) in vitro , we used Agrobacterium to introduce a CRISPR/Cas9 system into leaf tissue, and identified the presence of deletions in 27% of TcNPR3 copies in the treated tissues. The edited tissue exhibited an increased resistance to infection with the cacao pathogen Phytophthora tropicalis and elevated expression of downstream defense genes. Analysis of off-target mutagenesis in sequences similar to sgRNA target sites using high-throughput sequencing did not reveal mutations above background sequencing error rates. These results confirm the function of NPR3 as a repressor of the cacao immune system and demonstrate the application of CRISPR/Cas9 as a powerful functional genomics tool for cacao. Several stably transformed and genome edited somatic embryos were obtained via Agrobacterium -mediated transformation, and ongoing work will test the effectiveness of this approach at a whole plant level.

Transient Expression of CRISPR/Cas9 Machinery Targeting TcNPR3 Enhances Defense Response in Theobroma cacao

PubMed Central

Fister, Andrew S.; Landherr, Lena; Maximova, Siela N.; Guiltinan, Mark J.

2018-01-01

Theobroma cacao, the source of cocoa, suffers significant losses to a variety of pathogens resulting in reduced incomes for millions of farmers in developing countries. Development of disease resistant cacao varieties is an essential strategy to combat this threat, but is limited by sources of genetic resistance and the slow generation time of this tropical tree crop. In this study, we present the first application of genome editing technology in cacao, using Agrobacterium-mediated transient transformation to introduce CRISPR/Cas9 components into cacao leaves and cotyledon cells. As a first proof of concept, we targeted the cacao Non-Expressor of Pathogenesis-Related 3 (TcNPR3) gene, a suppressor of the defense response. After demonstrating activity of designed single-guide RNAs (sgRNA) in vitro, we used Agrobacterium to introduce a CRISPR/Cas9 system into leaf tissue, and identified the presence of deletions in 27% of TcNPR3 copies in the treated tissues. The edited tissue exhibited an increased resistance to infection with the cacao pathogen Phytophthora tropicalis and elevated expression of downstream defense genes. Analysis of off-target mutagenesis in sequences similar to sgRNA target sites using high-throughput sequencing did not reveal mutations above background sequencing error rates. These results confirm the function of NPR3 as a repressor of the cacao immune system and demonstrate the application of CRISPR/Cas9 as a powerful functional genomics tool for cacao. Several stably transformed and genome edited somatic embryos were obtained via Agrobacterium-mediated transformation, and ongoing work will test the effectiveness of this approach at a whole plant level. PMID:29552023

Immunomodulatory properties of cacao extracts – potential consequences for medical applications

PubMed Central

Becker, Kathrin; Geisler, Simon; Ueberall, Florian; Fuchs, Dietmar; Gostner, Johanna M.

2013-01-01

Anti-inflammatory properties of cacao, fruits of Theobroma cacao L. (Sterculiaceae), are well documented, and therapeutic applications are described for gastrointestinal, nervous, and cardiovascular abnormalities. Most, if not all of these disease conditions involve inflammation or immune activation processes. The pro-inflammatory cytokine interferon-γ (IFN-γ) and related biochemical pathways like tryptophan breakdown by indoleamine 2,3-dioxygenase (IDO) and neopterin formation are deeply involved in their pathogenesis. Neopterin concentrations and the kynurenine to tryptophan ratio (Kyn/Trp, an estimate of IDO activity) are elevated in a significant proportion of patients with virus infections, cancer, autoimmune syndrome, neurodegeneration, and coronary artery disease. Moreover, higher neopterin and Kyn/Trp concentrations are indicative for poor prognosis. When investigating the effect of aqueous or ethanolic extracts of cacao on IFN-γ, neopterin and Kyn/Trp concentrations in mitogen-stimulated human peripheral blood mononuclear cells, breakdown of tryptophan by IDO, and formation of neopterin and IFN-γ were dose-dependently suppressed. The effects observed in the cell-based assays are associated with the antioxidant activity of the cacao extracts as determined by the cell-free oxygen radical absorption capacity assay. The influence of cacao extracts on IDO activity could be of particular relevance for some of the beneficial health effects ascribed to cacao: tryptophan breakdown by IDO is strongly involved in immunoregulation, and the diminished availability of tryptophan limits the biosynthesis of neurotransmitter serotonin. The inhibition of tryptophan breakdown by cacao constituents could thus be relevant not only for immune system restoration in patients, but also contribute to mood elevation and thereby improve quality of life. However, the available data thus far are merely in vitro only and future studies need to investigate the influence of cacao on tryptophan metabolism in vivo. PMID:24376420

Characterization of Pseudomonas chlororaphis from Theobroma cacao L. rhizosphere with antagonistic activity against Phytophthora palmivora (Butler).

PubMed

Acebo-Guerrero, Y; Hernández-Rodríguez, A; Vandeputte, O; Miguélez-Sierra, Y; Heydrich-Pérez, M; Ye, L; Cornelis, P; Bertin, P; El Jaziri, M

2015-10-01

To isolate and characterize rhizobacteria from Theobroma cacao with antagonistic activity against Phytophthora palmivora, the causal agent of the black pod rot, which is one of the most important diseases of T. cacao. Among 127 rhizobacteria isolated from cacao rhizosphere, three isolates (CP07, CP24 and CP30) identified as Pseudomonas chlororaphis, showed in vitro antagonistic activity against P. palmivora. Direct antagonism tested in cacao detached leaves revealed that the isolated rhizobacteria were able to reduce symptom severity upon infection with P. palmivora Mab1, with Ps. chlororaphis CP07 standing out as a potential biocontrol agent. Besides, reduced symptom severity on leaves was also observed in planta where cacao root system was pretreated with the isolated rhizobacteria followed by leaf infection with P. palmivora Mab1. The production of lytic enzymes, siderophores, biosurfactants and HCN, as well as the detection of genes encoding antibiotics, the formation of biofilm, and bacterial motility were also assessed for all three rhizobacterial strains. By using a mutant impaired in viscosin production, derived from CP07, it was found that this particular biosurfactant turned out to be crucial for both motility and biofilm formation, but not for the in vitro antagonism against Phytophthora, although it may contribute to the bioprotection of T. cacao. In the rhizosphere of T. cacao, there are rhizobacteria, such as Ps. chlororaphis, able to protect plants against P. palmivora. This study provides a theoretical basis for the potential use of Ps. chlororaphis CP07 as a biocontrol agent for the protection of cacao plants from P. palmivora infection. © 2015 The Society for Applied Microbiology.

Cacao use and the San Lorenzo Olmec

PubMed Central

Powis, Terry G.; Cyphers, Ann; Gaikwad, Nilesh W.; Grivetti, Louis; Cheong, Kong

2011-01-01

Mesoamerican peoples had a long history of cacao use—spanning more than 34 centuries—as confirmed by previous identification of cacao residues on archaeological pottery from Paso de la Amada on the Pacific Coast and the Olmec site of El Manatí on the Gulf Coast. Until now, comparable evidence from San Lorenzo, the premier Olmec capital, was lacking. The present study of theobromine residues confirms the continuous presence and use of cacao products at San Lorenzo between 1800 and 1000 BCE, and documents assorted vessels forms used in its preparation and consumption. One elite context reveals cacao use as part of a mortuary ritual for sacrificial victims, an event that occurred during the height of San Lorenzo's power. PMID:21555564

Design and implementation of the cacao genome database

USDA-ARS?s Scientific Manuscript database

The Cacao Genome Database (CGD, www.cacaogenomedb.org) is being developed to provide a comprehensive data mining resource of genomic, genetic and breeding data for Theobroma cacao. Designed using Chado and a collection of Drupal modules, known as Tripal, CGD currently contains the genetically anchor...

Yield performance of cacao propagated by somatic embryogenesis and grafting

USDA-ARS?s Scientific Manuscript database

Twelve cacao (Theobroma cacao) clones propagated by grafting and somatic embryogenesis and grown on an Ultisol soil were evaluated for five years under intensive management at Corozal, Puerto Rico. Preliminary data showed no significant differences between propagation methods for yield of dry beans ...

Iron sources effects on growth, physiological parameters and nutrition of cacao

USDA-ARS?s Scientific Manuscript database

Productivity and sustainability of cacao (Theobroma cacao L.) in tropical soils are affected by deficiency of micronutrients. Iron deficiency is one of the main yield limiting constraints, especially in highly weathered, coarse textured and leached soils. To correct iron deficiency, different form...

Analyses of polyphenols in cacao liquor, cocoa, and chocolate by normal-phase and reversed-phase HPLC.

PubMed

Natsume, M; Osakabe, N; Yamagishi, M; Takizawa, T; Nakamura, T; Miyatake, H; Hatano, T; Yoshida, T

2000-12-01

The antioxidant polyphenols in cacao liquor, a major ingredient of chocolate and cocoa, have been characterized as flavan-3-ols and proanthocyanidin oligomers. In this study, various cacao products were analyzed by normal-phase HPLC, and the profiles and quantities of the polyphenols present, grouped by molecular size (monomers to approximately oligomers), were compared. Individual cacao polyphenols, flavan-3-ols (catechin and epicatechin), and dimeric (procyanidin B2), trimeric (procyanidin C1), and tetrameric (cinnamtannin A2) proanthocyanidins, and galactopyranosyl-ent-(-)-epicatechin (2alpha-->7, 4alpha-->8)-(-)-epicatechin (Gal-EC-EC), were analyzed by reversed-phase HPLC and/or HPLC/MS. The profile of monomers (catechins) and proanthocyanidin in dark chocolate was similar to that of cacao liquor, while the ratio of flavan-3-ols to the total amount of monomeric and oligomeric polyphenols in the case of pure cocoa powder was higher than that in the case of cacao liquor or chocolate.

Geographic and Genetic Population Differentiation of the Amazonian Chocolate Tree (Theobroma cacao L)

PubMed Central

Motamayor, Juan C.; Lachenaud, Philippe; da Silva e Mota, Jay Wallace; Loor, Rey; Kuhn, David N.; Brown, J. Steven; Schnell, Raymond J.

2008-01-01

Numerous collecting expeditions of Theobroma cacao L. germplasm have been undertaken in Latin-America. However, most of this germplasm has not contributed to cacao improvement because its relationship to cultivated selections was poorly understood. Germplasm labeling errors have impeded breeding and confounded the interpretation of diversity analyses. To improve the understanding of the origin, classification, and population differentiation within the species, 1241 accessions covering a large geographic sampling were genotyped with 106 microsatellite markers. After discarding mislabeled samples, 10 genetic clusters, as opposed to the two genetic groups traditionally recognized within T. cacao, were found by applying Bayesian statistics. This leads us to propose a new classification of the cacao germplasm that will enhance its management. The results also provide new insights into the diversification of Amazon species in general, with the pattern of differentiation of the populations studied supporting the palaeoarches hypothesis of species diversification. The origin of the traditional cacao cultivars is also enlightened in this study. PMID:18827930

Geographic and genetic population differentiation of the Amazonian chocolate tree (Theobroma cacao L).

PubMed

Motamayor, Juan C; Lachenaud, Philippe; da Silva E Mota, Jay Wallace; Loor, Rey; Kuhn, David N; Brown, J Steven; Schnell, Raymond J

2008-10-01

Numerous collecting expeditions of Theobroma cacao L. germplasm have been undertaken in Latin-America. However, most of this germplasm has not contributed to cacao improvement because its relationship to cultivated selections was poorly understood. Germplasm labeling errors have impeded breeding and confounded the interpretation of diversity analyses. To improve the understanding of the origin, classification, and population differentiation within the species, 1241 accessions covering a large geographic sampling were genotyped with 106 microsatellite markers. After discarding mislabeled samples, 10 genetic clusters, as opposed to the two genetic groups traditionally recognized within T. cacao, were found by applying Bayesian statistics. This leads us to propose a new classification of the cacao germplasm that will enhance its management. The results also provide new insights into the diversification of Amazon species in general, with the pattern of differentiation of the populations studied supporting the palaeoarches hypothesis of species diversification. The origin of the traditional cacao cultivars is also enlightened in this study.

Anti-HIV and immunomodulation activities of cacao mass lignin-carbohydrate complex.

PubMed

Sakagami, Hiroshi; Kawano, Michiyo; Thet, May Maw; Hashimoto, Ken; Satoh, Kazue; Kanamoto, Taisei; Terakubo, Shigemi; Nakashima, Hideki; Haishima, Yuji; Maeda, Yuuichi; Sakurai, Koji

2011-01-01

Recently, a prominent antiviral and macrophage stimulatory activity of cacao lignin-carbohydrate complex (LCC) has been reported. However, the solubility and sterility of LCC have not been considered yet. In the present study, complete solubilisation and sterilisation was achieved by autoclaving under mild alkaline conditions and the previously reported biological activities were re-examined. LCCs were obtained by 1% NaOH extraction and acid precipitation, and a repeated extraction-precipitation cycle. Nitric oxide (NO) and cytokine productions were assayed by the Griess method and ELISA, respectively. Inducible NO synthase (iNOS) expression was determined by Western blot analysis. Superoxide anion, hydroxyl radical and nitric oxide radical-scavenging activity was determined by ESR spectroscopy. Cacao mass LCC showed reproducibly higher anti-HIV activity than cacao husk LCC. Cacao mass LCC, up to 62.5 μg/ml, did not stimulate mouse macrophage-like cells (RAW264.7 and J774.1) to produce NO, nor did it induce iNOS protein, in contrast to lipopolysaccharide (LPS). Cacao mass LCC and LPS synergistically stimulated iNOS protein expression, suggesting a different point of action. Cacao mass LCC induced tumour necrosis factor-α production markedly less than LPS, and did not induce interleukin-1β, interferon-α or interferon-γ. ESR spectroscopy showed that cacao mass LCC, but not LPS, scavenged NO produced from NOC-7. This study demonstrated several new biological activities of LCCs distinct from LPS and further confirmed the promising antiviral and immunomodulating activities of LCCs.

Fluorescent detection of (-)-epicatechin in microsamples from cacao seeds and cocoa products: Comparison with Folin-Ciocalteu method.

PubMed

Ramirez-Sanchez, Israel; Maya, Lisandro; Ceballos, Guillermo; Villarreal, Francisco

2010-12-01

Polyphenolic compounds of the flavanoid family are abundantly present in cacao seed and its cocoa products. Results from studies using cocoa products indicate beneficial effects of flavanols on cardiovascular endpoints. Evidence indicates that (-)-epicatechin is the main cacao flavanol associated with cardiovascular effects, so the accurate quantification of its content in cacao seeds or cocoa products is important. Common methods for the quantification of phenolic content in cocoa products are based on the reaction of phenols with colorimetric reagents such as the Folin-Ciocalteu (FC) In this study, we compared the FC method of phenolic determinations using 2 different standards (gallic acid and (-)-epicatechin) to construct calibration curves. We compare these results with those obtained from a simple fluorometric method (Ex(280)/Em(320) nm) used to determine catechin/(-)-epicatechin content in samples of cacao seeds and cocoa products. Values obtained from the FC method determination of polyphenols yield an overestimation of phenol (flavonoid) content when gallic acid is used as standard. Moreover, the epicatechin is a more reliable standard because of its abundance in cacao seeds and cocoa products. The use of fluorometric spectra yields a simple and highly quantitative means for a more precise and rapid quantification of cacao catechins. Fluorometric values are essentially in agreement with those reported using more cumbersome methods. In conclusion, the use of fluorescence emission spectra is a quick, practical and suitable means to quantifying catechins in cacao seeds and cocoa products.

Genetic diversity of naturalized cacao (Theobroma cacao L.) in Puerto Rico

USDA-ARS?s Scientific Manuscript database

Identification of genetically diverse cacao with disease resistance, high productivity and desirable organoleptic traits is vitally important to the agricultural crop’s long-term sustainability. Environmental changes, pests and diseases as well as nation’s sovereign property rights have led to a de...

Origin, dispersal and current global distribution of cacao genetic diversity

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.) is cultivated globally as the unique source of cocoa butter and powder for the confectionery industries. In spite of its economical importance, cocoa was and continues to be dominantly produced in low-input and low-output systems. Production constraints, including depletio...

A genetically anchored physical map of the cacao genome

USDA-ARS?s Scientific Manuscript database

Mars Incorporated and the United States Department of Agriculture have undertaken the sequencing of the genome of Theobroma cacao, which produces cocoa beans, the key ingredient in chocolate. Genetic information, such as whole genome sequence is necessary to better understand and improve cacao. In m...

The Mitochondrial Genome of Moniliophthora roreri, the frosty pod rot pathogen of cacao

USDA-ARS?s Scientific Manuscript database

Moniliophthora roreri and Moniliophthora perniciosa are closely related basidiomycetes that cause two important diseases in cacao (Theobroma cacao L.): frosty pod rot and the witches' broom disease, respectively. A comparison of the complete mitochondrial genomes of these pathogens shows a high degr...

Cacao genetic resources research at the USDA-ARS Tropical Agriculture Research Station, Mayaguez, Puerto Rico

USDA-ARS?s Scientific Manuscript database

The current USDA ARS Tropical Agriculture Research Station’s (TARS) cacao (Theobroma cacao L.) collection in Mayaguez, Puerto Rico, consists of 262 clonally propagated accessions. Each accession is represented by six individual trees grafted onto a common ‘Amelonado’ seedling rootstock and planted ...

Association mapping of fruit, seed and disease resistance traits in Theobroma cacao L

USDA-ARS?s Scientific Manuscript database

An association mapping approach was employed to find markers for color, size, girth and mass of fruits; seed number and butterfat content; and resistance to black pod and witches’ broom diseases in cacao (Theobroma cacao L.). Ninety-five microsatellites (SSRs) and 775 single nucleotide polymorphisms...

Moniliophthora roreri Genome and Transcriptome

USDA-ARS?s Scientific Manuscript database

Frosty pod rot disease of cacao is one of the most destructive diseases of cacao and at this time is limited to regions in South America and Central America. Frosty pod rot is caused by a fungal pathogen Moniliophthora roreri, a basidiomycete that is closely related to another cacao pathogen that ca...

Sequencing the Cacao Genome: Overall Strategy and SNP Discovery for Cacao Improvement.

USDA-ARS?s Scientific Manuscript database

On June 26, 2008, the United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Mars, Incorporated, and IBM announced that they are combining their scientific resources to sequence and analyze the entire genome of Theobroma cacao L., an understory tree from the Amazon basin w...

Molecular, physiological and morphological analysis of waterlogging tolerance in clonal genotypes of Theobroma cacao

USDA-ARS?s Scientific Manuscript database

In soil, hypoxia and anoxia conditions generated by waterlogging induce changes in genetic morphological, physiological processes, and as well as altering the growth and development of plant The mass propagation of cacao (Theobroma cacao) cuttings-to produce plantlets (clones) is affected by waterlo...

A genome survey of Moniliophthora perniciosa gives new insights into Witches’ Broom Disease of cacao

USDA-ARS?s Scientific Manuscript database

Background: The basidiomycete fungus Moniliophthora perniciosa is the causal agent of Witches’ Broom Disease (WBD) in cacao (Theobroma cacao). It is a hemibiotrophic pathogen that colonizes the apoplast of cacao’s meristematic tissues as a biotrophic pathogen, switching to a saprotrophic lifestyle d...

Transcriptome characterization for genome annotation and functional genomics in Theobroma cacao

USDA-ARS?s Scientific Manuscript database

Evidence from leaf transcriptome sequencing using two technology platforms, in combination with protein homology and trained ab initio predictions, previously enabled us to build 35,000 gene models in T. cacao (www.cacaogenomedb.org). Here we review the contribution of each data type to cacao gene a...

Influence of low light intensity and soil flooding on cacao physiology

USDA-ARS?s Scientific Manuscript database

Growth and development of plants frequently are limited by multiple abiotic stresses that occur simultaneously in the environment. Cabruca’ an agroforestry system is a main cropping system invariably adapted for cultivation of cacao in southern Bahia, Brazil. In this system of management cacao is gr...

Factors influencing the survival of developing embryos of theobroma cacao L. (Malvaceae) in cryogenic storage

USDA-ARS?s Scientific Manuscript database

Cacao, Theobroma cacao L., is native to tropical South American rainforests and is the source of chocolate. Ex situ conservation of this economically important species and its relatives is imperative to prevent genetic erosion resulting from diminished suitable habitat and increased pressure from a...

Antimicrobial and plant growth-promoting properties of the cacao endophyte Bacillus subtilis ALB629.

PubMed

Falcäo, L L; Silva-Werneck, J O; Vilarinho, B R; da Silva, J P; Pomella, A W V; Marcellino, L H

2014-06-01

To investigate the effects of the endophyte Bacillus subtilisALB629 on the growth of cacao seedlings at early developmental stage and to evaluate its antimicrobial properties. Germinating cacao seeds were inoculated with ALB629, and seedlings growth was evaluated 30 days later. Significant increase (P < 0·05) was observed in the root system (up to 30%), leaf area (14%) and stem height (7·6%). ALB629 colonized the entire plant, prevailing over indigenous micro-organisms. In addition, it was tested in vitro, by pairing assays, and showed antagonistic effect against the phytopathogenic fungi Moniliophthora perniciosa, Colletotrichum sp. and C. gossypii. When tested in cacao-grafting procedure in the field, ALB629 increased the grafting success rate (24%), indicating its protective effect. In addition, this Bacillus secretes an antagonist compound, as shown by the antifungal activity of the cell-free culture. Bacillus subtilisALB629 promotes cacao root growth, besides promoting growth of the aerial part of cacao seedlings. It has antimicrobial properties and produces an antifungal compound. ALB629 presented beneficial characteristics for cacao cultivation, being a good biological control agent candidate. Furthermore, it is a potential source of antifungal compound with potential for commercial exploitation. © 2014 The Society for Applied Microbiology.

Cacao Phylloplane: The First Battlefield against Moniliophthora perniciosa, Which Causes Witches' Broom Disease.

PubMed

Almeida, D S M; Gramacho, K P; Cardoso, T H S; Micheli, F; Alvim, F C; Pirovani, C P

2017-07-01

The phylloplane is the first contact surface between Theobroma cacao and the fungus Moniliophthora perniciosa, which causes witches' broom disease (WBD). We evaluated the index of short glandular trichomes (SGT) in the cacao phylloplane and the effect of irrigation on the disease index of cacao genotypes with or without resistance to WBD, and identified proteins present in the phylloplane. The resistant genotype CCN51 and susceptible Catongo presented a mean index of 1,600 and 700 SGT cm -2 , respectively. The disease index in plants under drip irrigation was reduced by approximately 30% compared with plants under sprinkler irrigation prior to inoculation. Leaf water wash (LWW) of the cacao inhibited the germination of spores by up to 98%. Proteins from the LWW of CCN51 were analyzed by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by tandem mass spectrometry. The gel showed 71 spots and identified a total of 42 proteins (28 from the plant and 14 from bacteria). Proteins related to defense and synthesis of defense metabolites and involved in nucleic acid metabolism were identified. The results support the hypothesis that the proteins and water-soluble compounds secreted to the cacao phylloplane participate in the defense against pathogens. They also suggest that SGT can contribute to the resistance of cacao.

Tree spacing impacts the individual incidence of Moniliophthora roreri disease in cacao agroforests.

PubMed

Ngo Bieng, Marie Ange; Alem, Laudine; Curtet, Chloé; Tixier, Philippe

2017-11-01

Using conventional pesticides in crop protection has raised serious environmental concerns and there is therefore a need for integrated pest management (IPM) methods. In this paper, we found that the spacing of trees can impact disease, which could result in a reduction in pesticide applications and may act as a potential IPM method. We studied Frosty Pod Rot (FPR) in 20 cacao agroforests in Costa Rica (Upala region). Using a generalized linear mixed model, we analyzed the impact of the neighborhood composition and distance from a studied cacao individual on its individual FPR incidence. We found that the number of cacao tree neighbors in a radius of 3.7 m and the number of fruit trees in a radius of 4.3 m had a significant negative influence on the incidence of FPR on individual cacao trees. Moreover, cacao tree neighbors had the most significant local influence compared to the neighborhood of other taller categories such as fruit or forest trees. The mechanisms involved are related to the barrier effect, due to the effectiveness of the cacao tree's architecture as an efficient barrier against FPR spore dispersal. This paper provides new insights into optimization of the spatial environment around each host as an original IPM method. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Diversity of endophytic fungal community of cacao (Theobroma cacao L.) and biological control of Crinipellis perniciosa, causal agent of Witches' Broom Disease.

PubMed

Rubini, Marciano R; Silva-Ribeiro, Rute T; Pomella, Alan W V; Maki, Cristina S; Araújo, Welington L; Dos Santos, Deise R; Azevedo, João L

2005-01-01

The basidiomycete fungus Crinipellis perniciosa (Stahel) Singer is the causal agent of Witches' Broom Disease of Cacao (Theobromacacao L.) which is the main factor limiting cacao production in the Americas. Pod losses of up to 90% are experienced in affected areas as evidenced by the 50% drop in production in Bahia province, Brazil following the arrival of the C. perniciosa in the area in 1989. The disease has proven particularly difficult to control and many farmers in affected areas have given up cacao cultivation. In order to evaluate the potential of endophytes as a biological control agent of this phytopathogen, the endophytic fungal community of resistant and susceptible cacao plants as well as affected branches was studied between 2001 and 2002. The fungal community was identified by morphological traits and rDNA sequencing as belonging to the genera Acremonium, Blastomyces, Botryosphaeria, Cladosporium, Colletotrichum, Cordyceps, Diaporthe, Fusarium, Geotrichum, Gibberella, Gliocladium, Lasiodiplodia, Monilochoetes, Nectria, Pestalotiopsis, Phomopsis, Pleurotus, Pseudofusarium, Rhizopycnis, Syncephalastrum, Trichoderma, Verticillium and Xylaria. These fungi were evaluated both in vitro and in vivo by their ability to inhibit C. perniciosa. Among these, some were identified as potential antagonists, but only one fungus (Gliocladium catenulatum) reduced the incidence of Witches' Broom Disease in cacao seedlings to 70%.

Management of Chinese Rose Beetle (Adoretus sinicus) Adults Feeding on Cacao (Theobroma cacao) Using Insecticides

PubMed Central

Spafford, Helen; Ching, Alexander; Manley, Megan; Hardin, Chelsea; Bittenbender, Harry

2016-01-01

The Chinese rose beetle (Adoretus sinicus Burmeister (Coleoptera: Scarabaeidae)) is an introduced, widely-established pest in Hawai’i. The adult beetles feed on the leaves of cacao (Theobroma cacao L.), which can lead to defoliation and even death of young trees. We evaluated the impact of five commercially available products with different active ingredients (imidacloprid, azadirachtin, Beauveria bassiana (Bals.-Criv.) Vuill., kaolin clay, and pyrethrin) and the presence or absence of weed mat cover in reducing adult beetle feeding on sapling cacao in the field. The use of weed mat cover reduced feeding damage compared to the untreated control, as did foliar application of imidacloprid, azadirachtin, and B. bassiana. In the laboratory, field-collected adult beetles were presented cacao leaf samples dipped in one of the five products and compared to a control. Beetles exposed to pyrethrin died rapidly. Among the other treatments, only exposure to imidacloprid significantly reduced survival relative to the control. Beetles fed very little on leaf samples with azadirachtin but their longevity was not significantly reduced. Imidacloprid, azadirachtin, and weed mat application had the most promise for reducing adult Chinese rose beetle feeding damage in young cacao and deserve further investigation for successful management of this significant pest. PMID:27348004

Vascular Streak Dieback of cacao in Southeast Asia and Melanesia: in planta detection of the pathogen and a new taxonomy.

PubMed

Samuels, Gary J; Ismaiel, Adnan; Rosmana, Ade; Junaid, Muhammad; Guest, David; McMahon, Peter; Keane, Philip; Purwantara, Agus; Lambert, Smilja; Rodriguez-Carres, Marianela; Cubeta, Marc A

2012-01-01

Vascular Streak Dieback (VSD) disease of cacao (Theobroma cacao) in Southeast Asia and Melanesia is caused by a basidiomycete (Ceratobasidiales) fungus Oncobasidium theobromae (syn. =Thanatephorus theobromae). The most characteristic symptoms of the disease are green-spotted leaf chlorosis or, commonly since about 2004, necrotic blotches, followed by senescence of leaves beginning on the second or third flush behind the shoot apex, and blackening of infected xylem in the vascular traces at the leaf scars resulting from the abscission of infected leaves. Eventually the shoot apex is killed and infected branches die. In susceptible cacao the fungus may grow through the xylem down into the main stem and kill a mature cacao tree. Infections in the stem of young plants prior to the formation of the first 3-4 lateral branches usually kill the plant. Basidiospores released from corticioid basidiomata developed on leaf scars or along cracks in the main vein of infected leaves infect young leaves. The pathogen commonly infects cacao but there are rare reports from avocado. As both crops are introduced to the region, the pathogen is suspected to occur asymptomatically in native vegetation. The pathogen is readily isolated but cultures cannot be maintained. In this study, DNA was extracted from pure cultures of O. theobromae obtained from infected cacao plants sampled from Indonesia. The internal transcribed spacer region (ITS), consisting of ITS1, 5.8S ribosomal RNA and ITS2, and a portion of nuclear large subunit (LSU) were sequenced. Phylogenetic analysis of ITS sequences placed O. theobromae sister to Ceratobasidium anastomosis groups AG-A, AG-Bo, and AG-K with high posterior probability. Therefore the new combination Ceratobasidium theobromae is proposed. A PCR-based protocol was developed to detect and identify C. theobromae in plant tissue of cacao enabling early detection of the pathogen in plants. A second species of Ceratobasidium, Ceratobasidium ramicola, identified through ITS sequence analysis, was isolated from VSD-affected cacao plants in Java, and is widespread in diseased cacao collected from Indonesia. Published by Elsevier Ltd.

Dispersion models and sampling of cacao mirid bug Sahlbergella singularis (Hemiptera: Miridae) on Theobroma Cacao in southern Cameroon.

PubMed

Bisseleua, D H B; Vidal, Stefan

2011-02-01

The spatio-temporal distribution of Sahlbergella singularis Haglung, a major pest of cacao trees (Theobroma cacao) (Malvaceae), was studied for 2 yr in traditional cacao forest gardens in the humid forest area of southern Cameroon. The first objective was to analyze the dispersion of this insect on cacao trees. The second objective was to develop sampling plans based on fixed levels of precision for estimating S. singularis populations. The following models were used to analyze the data: Taylor's power law, Iwao's patchiness regression, the Nachman model, and the negative binomial distribution. Our results document that Taylor's power law was a better fit for the data than the Iwao and Nachman models. Taylor's b and Iwao's β were both significantly >1, indicating that S. singularis aggregated on specific trees. This result was further supported by the calculated common k of 1.75444. Iwao's α was significantly <0, indicating that the basic distribution component of S. singularis was the individual insect. Comparison of negative binomial (NBD) and Nachman models indicated that the NBD model was appropriate for studying S. singularis distribution. Optimal sample sizes for fixed precision levels of 0.10, 0.15, and 0.25 were estimated with Taylor's regression coefficients. Required sample sizes increased dramatically with increasing levels of precision. This is the first study on S. singularis dispersion in cacao plantations. Sampling plans, presented here, should be a tool for research on population dynamics and pest management decisions of mirid bugs on cacao. © 2011 Entomological Society of America

Chemical speciation of cadmium: an approach to evaluate plant-available cadmium in Ecuadorian soils under cacao production

USDA-ARS?s Scientific Manuscript database

Elevated concentration of cadmium (Cd) in cacao beans has raised serious concerns about the safety of chocolate consumption. Accumulation of Cd cacao bean in southern Ecuador has been reported to relate soil contamination. In this study, soil fractionation was conducted to identify available Cd poo...

21 CFR 163.5 - Methods of analysis.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Methods of analysis. 163.5 Section 163.5 Food and... CONSUMPTION CACAO PRODUCTS General Provisions § 163.5 Methods of analysis. Shell and cacao fat content in cacao products shall be determined by the following methods of analysis prescribed in “Official Methods...

First report of frosty pod rot caused by Moniliophthora roreri on cacao in Bolivia

USDA-ARS?s Scientific Manuscript database

Frosty pod rot (FPR) is a devastating cacao disease caused by the basidiomycete Moniliophthora roreri (Aime and Phillips-Mora, 2005). The disease is confined to 13 countries in Central and South America and constitutes a permanent threat for cacao cultivation worldwide. In July 2012, FPR was detect...

Diallel analysis and growth parameters as selection tools for drought tolerance in young Theobroma cacao plants

USDA-ARS?s Scientific Manuscript database

Technical Abstract: This study was aimed to estimate the combining ability, through diallel crosses, of T. cacao genotypes preselected for drought tolerance. The experiment was conducted under greenhouse conditions at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a completely randomiz...

Making a chocolate chip: development and evaluation of a 6K SNP array for Theobroma cacao.

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the key ingredient in chocolate production, is one of the world's most important tree fruit crops, with ~4,000,000 metric tons produced across 50 countries. To move towards gene discovery and marker-assisted breeding in cacao, a single-nucleotide polymorphism (SNP) identification pr...

Toward The Identification Of Candidate Genes Involved In Black Pod Disease Resistance In Theobroma cacao L.

USDA-ARS?s Scientific Manuscript database

Black Pod (Phytophthora sp.) has a devastating effect on the worldwide cacao (Theobroma cacao) yield and incorporating resistance into production fields has been an ongoing effort of breeding programs. Previous meta-QTL analysis of genetic maps created with a variety of molecular markers identified...

Yield performance and bean quality traits of cacao propagated by somatic embryogenesis and grafting

USDA-ARS?s Scientific Manuscript database

Twelve cacao (Theobroma cacao) clones propagated by grafting and rooted cuttings of somatic embryo-derived plants were grown on an Ultisol soil at Corozal, Puerto Rico and evaluated for six years under intensive management. Year, variety, the year x variety and propagation treatment x variety intera...

Toward the identification of candidate genes involved In black pod disease resistance in Theobroma cacao L.

USDA-ARS?s Scientific Manuscript database

Black Pod (Phytophthora sp.) has a devastating effect on the worldwide cacao (Theobroma cacao) yield and incorporating resistance into production fields has been an ongoing effort of breeding programs. Previous meta-QTL analysis of genetic maps created with a variety of molecular markers identified...

Vascular Streak Dieback of cacao in Southeast Asia detection and Melanesia: in planta detection of the pathogen and a new taxonomy

USDA-ARS?s Scientific Manuscript database

Vascular Streak Dieback (VSD) disease of cacao (Theobroma cacao) in Southeast Asia and Melanesia is caused by a basidiomycete (Ceratobasidiales) fungus described in a monotypic genus as Oncobasidium theobromae (syn. =Thanatephorus theobromae). The symptoms of the disease include green-spotted chloro...

Yield performance and bean quality traits of cacao propagated by grafting and somatic embryo-derived cuttings

USDA-ARS?s Scientific Manuscript database

Twelve cacao (Theobroma cacao) clones propagated by grafting and orthotropic rooted cuttings of somatic embryo-derived plants were grown on an Ultisol soil at Corozal, Puerto Rico and evaluated for six years of production under intensive management. Year, variety, year x variety and propagation tre...

Aspergillus oryzae NRRL 35191 from coffee, a non-toxigenic endophyte with the ability to synthesize kojic acid

USDA-ARS?s Scientific Manuscript database

Aspergillus oryzae was isolated as an endophyte from coffee leaves and found to produce kojic acid in culture. When inoculated in cacao seedlings (Theobroma cacao L.), A. oryzae grew endophytically and synthesize kojic acid in planta. Cacao seedlings inoculated with A. oryzae produced higher levels...

Nuclear and mitochondrial rDNA variability in Crinipellis perniciosa from different geographic origins and hosts.

PubMed

de Arruda, Maricília C C; Ferreira, Marisa A S V; Miller, Robert N G; Resende, Mário Lúcio V; Felipe, Maria Sueli S

2003-01-01

Genetic variability in Crinipellis perniciosa, the causal organism of witches' broom disease in Theobroma cacao, was determined in strains originating from T. cacao and other susceptible host species Heteropterys acutifolia and Solanum lycocarpum in Brazil, in order to clarify host specificity and geographical variability. RFLP analysis of the ribosomal DNA ITS regions (rDNA ITS), and the mitochondrial DNA small subunit ribosomal DNA gene (mtDNA SSU rDNA) did not reveal any genetic variability in 120 tested strains, possibly serving only as species level markers. Genetic variability was observed in the ribosomal DNA IGS spacer region, in terms of IGS size, RFLPs and sequence data. Phylogenetic analyses (using CLUSTAL W, PHYLIP and TREEVIEW) indicated considerable differences between C. perniciosa strains from T. cacao and those from H. acutifolia (85-86%) and S. lycocarpum (95-96%). Sequence differences also indicated that C. perniciosa from T. cacao in Bahia is less variable (98%) when compared to the pathogen on T. cacao in Amazonas (97-98%), perhaps reflecting a recent introduction to T. cacao in Bahia.

STATUS OF CACAO WITCHES' BROOM: biology, epidemiology, and management.

PubMed

Purdy, L H; Schmidt, R A

1996-01-01

Origins of Theobroma cacao and Crinipellis perniciosa occurred in the Amazon Basin region of South America, and their interaction, the witches' broom disease, was first described in the late 1700s. The 100 years of scientific investigations of witches' broom of cacao that began in the 1890s developed the present state of knowledge of the biology and epidemiology of witches' broom that are discussed. Recommended management to reduce the deleterious effects of witches' broom on cacao production include the use of phytosanitation (removal of diseased plant parts), applications of chemical fungicides, and the use of host resistance. At present, there is a paucity of resistant planting materials, and efforts to evaluate germplasm for resistance to witches' broom are described. Research topics to augment present knowledge about witches' broom of cacao are presented with the hope that disease management can be improved.

Endophytic fungal diversity in Theobroma cacao (cacao) and T. grandiflorum (cupuaçu) trees and their potential for growth promotion and biocontrol of black-pod disease.

PubMed

Hanada, Rogério Eiji; Pomella, Alan William V; Costa, Heron Salazar; Bezerra, José Luiz; Loguercio, Leandro L; Pereira, José O

2010-01-01

The endophytic niches of plants are a rich source of microbes that can directly and indirectly promote plant protection, growth and development. The diversity of culturable endophytic fungi from stems and branches of Theobroma cacao (cacao) and Theobroma grandiflorum (cupuaçu) trees growing in the Amazon region of Brazil was assessed. The collection of fungal endophytic isolates obtained was applied in field experiments to evaluate their potential as biocontrol agents against Phytophthora palmivora, the causal agent of the black-pod rot disease of cacao, one of the most important pathogens in cocoa-producing regions worldwide. The isolated endophytic fungi from 60 traditional, farmer-planted, healthy cacao and 10 cupuaçu plants were cultured in PDA under conditions inducing sporulation. Isolates were classified based upon the morphological characteristics of their cultures and reproductive structures. Spore suspensions from a total of 103 isolates that could be classified at least up to genus level were tested against P. palmivora in pods attached to cacao trees in the field. Results indicated that ∼70% of isolates showed biocontrol effects to a certain extent, suggesting that culturable endophytic fungal biodiversity in this system is of a mostly mutualistic type of interaction with the host. Eight isolates from genera Trichoderma (reference isolate), Pestalotiopsis, Curvularia, Tolypocladium and Fusarium showed the highest level of activity against the pathogen, and were further characterized. All demonstrated their endophytic nature by colonizing axenic cacao plantlets, and confirmed their biocontrol activity on attached pods trials by showing significant decrease in disease severity in relation to the positive control. None, however, showed detectable growth-promotion effects. Aspects related to endophytic biodiversity and host-pathogen-endophyte interactions in the environment of this study were discussed on the context of developing sustainable strategies for biological control of black-pod rot of cacao. Copyright © 2010 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Suppressive effects of cacao polyphenols on the development of atherosclerosis in apolipoprotein E-deficient mice.

PubMed

Natsume, Midori; Baba, Seigo

2014-01-01

Previous studies in humans have shown that the cacao polyphenols, (-)-epicatechin and its oligomers, prevent in vitro and ex vivo low-density lipoprotein oxidation mediated by free radical generators and metal ions and also reduce plasma LDL-cholesterol levels. The aim of this study was to examine the effects of cacao polyphenols on the development of atherosclerosis in apolipoprotein E-deficient (-/-) mice. Mice aged 8 weeks (n = 90) were randomized into three groups, and fed either normal mouse chow (controls) or chow supplemented with 0.25 or 0.40 % cacao polyphenols for 16 weeks. The mean plaque area in cross-sections of the brachiocephalic trunk was measured and found to be lower in the 0.25 % cacao polyphenol group than in the control group (p < 0.05). Pathological observations showed that accumulation of cholesterol crystals in the plaque area was greater in the control group compared with the 0.40 % cacao polyphenol group (p < 0.05). Immunochemical staining in the 0.25 and 0.40 % groups showed that expression of the cell adhesion molecules (VCAM-1 and ICAM-1) and production of oxidative stress markers (4-hydroxynonenal, hexanoyl-lysine, and dityrosine) were reduced in cross-sections of the brachiocephalic trunk. These results suggest that cacao polyphenols inhibit the development of atherosclerosis in apolipoprotein E-deficient (-/-) mice by reducing oxidative stress and inflammatory responses.

Molecular characterization of previously elusive badnaviruses associated with symptomatic cacao in the New World.

PubMed

Chingandu, Nomatter; Zia-Ur-Rehman, Muhammad; Sreenivasan, Thyail N; Surujdeo-Maharaj, Surendra; Umaharan, Pathmanathan; Gutierrez, Osman A; Brown, Judith K

2017-05-01

Suspected virus-like symptoms were observed in cacao plants in Trinidad during 1943, and the viruses associated with these symptoms were designated as strains A and B of cacao Trinidad virus (CTV). However, viral etiology has not been demonstrated for either phenotype. Total DNA was isolated from symptomatic cacao leaves exhibiting the CTV A and B phenotypes and subjected to Illumina HiSeq and Sanger DNA sequencing. Based on de novo assembly, two apparently full-length badnavirus genomes of 7,533 and 7,454 nucleotides (nt) were associated with CTV strain A and B, respectively. The Trinidad badnaviral genomes contained four open reading frames, three of which are characteristic of other known badnaviruses, and a fourth that is present in only some badnaviruses. Both badnaviral genomes harbored hallmark caulimovirus-like features, including a tRNA Met priming site, a TATA box, and a polyadenylation-like signal. Pairwise comparisons of the RT-RNase H region indicated that the Trinidad isolates share 57-71% nt sequence identity with other known badnaviruses. Based on the system for badnavirus species demarcation in which viruses with less than 80% nt sequence identity in the RT-RNase gene are considered members of separate species, these isolates represent two previously unidentified badnaviruses, herein named cacao mild mosaic virus and cacao yellow vein banding virus, making them the first cacao-infecting badnaviruses identified thus far in the Western Hemisphere.

Genome Size Evolution in Theobroma cacao: Recent Sequencing of Two Cacao Genomes of Different Size

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the source of cocoa beans for chocolate, is an important tropical agriculture commodity that is affected by a number of fungal pathogens and insect pests, as well as concerns about yield and quality. We are trying to find molecular genetic markers that are linked to disease resista...

A SNP assay for genotyping Theobroma cacao suitable for use under field conditions

USDA-ARS?s Scientific Manuscript database

The tropical tree crop Theobroma cacao L. is grown commercially for its beans, which are used in the production of cocoa butter and chocolate. Although the upper Amazon region of South America is the center of origin for cacao, 70% of the world’s supply of cocoa beans currently comes from small far...

Yield performance and bean quality traits of cacao propagated by grafting and somatic embryo-derived cuttings

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao) has great potential as a component of a small tropical farming system. It adapts to a wide range of soils of climatic conditions, grows well under minimum tillage, adapts to temporary intercropping, has the potential of being sold in local and export markets and the pods are ...

El nombre 'Forastero' no más: A new protocol for meaningful cacao germplasm classification.

USDA-ARS?s Scientific Manuscript database

The title of this article (The name ‘Forastero’ no more) is to convey an attempt in this paper to try to convince the cacao scientific community not to use the term Forastero to identify cacao germplasm of non-Criollo origin. The term Forastero originated in Latin America to differentiate the intro...

Impact of drought on morphological, physiological and nutrient use efficiency of elite cacao genotypes from Bahia-Brazil, Tarapoto-Peru and Puerto Rico-USA

USDA-ARS?s Scientific Manuscript database

Worldwide, drought is considered one of the most limiting abiotic stress factors for cacao growth, development and production. A series of greenhouse and growth chamber experiments were undertaken to assess drought effects on early cacao morphological and physiological traits and nutrient use effici...

Impact of drought on morphological, physiological and nutrient use efficiency of elite Cacao genotypes from Bahia-Brazil, Tarapoto-Peru and Puerto Rico-USA

USDA-ARS?s Scientific Manuscript database

Worldwide, drought is considered one of the most limiting abiotic stress factors for cacao growth, development and production. Climatic changes that are occurring in the tropics such as inconsistent and reduced rainfall patterns, and high temperatures in many cacao growing regions has impacted yiel...

The genome sequence of the most widely cultivated cacao type and its use to identify candidate genes regulating pod color.

PubMed

Motamayor, Juan C; Mockaitis, Keithanne; Schmutz, Jeremy; Haiminen, Niina; Livingstone, Donald; Cornejo, Omar; Findley, Seth D; Zheng, Ping; Utro, Filippo; Royaert, Stefan; Saski, Christopher; Jenkins, Jerry; Podicheti, Ram; Zhao, Meixia; Scheffler, Brian E; Stack, Joseph C; Feltus, Frank A; Mustiga, Guiliana M; Amores, Freddy; Phillips, Wilbert; Marelli, Jean Philippe; May, Gregory D; Shapiro, Howard; Ma, Jianxin; Bustamante, Carlos D; Schnell, Raymond J; Main, Dorrie; Gilbert, Don; Parida, Laxmi; Kuhn, David N

2013-06-03

Theobroma cacao L. cultivar Matina 1-6 belongs to the most cultivated cacao type. The availability of its genome sequence and methods for identifying genes responsible for important cacao traits will aid cacao researchers and breeders. We describe the sequencing and assembly of the genome of Theobroma cacao L. cultivar Matina 1-6. The genome of the Matina 1-6 cultivar is 445 Mbp, which is significantly larger than a sequenced Criollo cultivar, and more typical of other cultivars. The chromosome-scale assembly, version 1.1, contains 711 scaffolds covering 346.0 Mbp, with a contig N50 of 84.4 kbp, a scaffold N50 of 34.4 Mbp, and an evidence-based gene set of 29,408 loci. Version 1.1 has 10x the scaffold N50 and 4x the contig N50 as Criollo, and includes 111 Mb more anchored sequence. The version 1.1 assembly has 4.4% gap sequence, while Criollo has 10.9%. Through a combination of haplotype, association mapping and gene expression analyses, we leverage this robust reference genome to identify a promising candidate gene responsible for pod color variation. We demonstrate that green/red pod color in cacao is likely regulated by the R2R3 MYB transcription factor TcMYB113, homologs of which determine pigmentation in Rosaceae, Solanaceae, and Brassicaceae. One SNP within the target site for a highly conserved trans-acting siRNA in dicots, found within TcMYB113, seems to affect transcript levels of this gene and therefore pod color variation. We report a high-quality sequence and annotation of Theobroma cacao L. and demonstrate its utility in identifying candidate genes regulating traits.

The genome sequence of the most widely cultivated cacao type and its use to identify candidate genes regulating pod color

PubMed Central

2013-01-01

Background Theobroma cacao L. cultivar Matina 1-6 belongs to the most cultivated cacao type. The availability of its genome sequence and methods for identifying genes responsible for important cacao traits will aid cacao researchers and breeders. Results We describe the sequencing and assembly of the genome of Theobroma cacao L. cultivar Matina 1-6. The genome of the Matina 1-6 cultivar is 445 Mbp, which is significantly larger than a sequenced Criollo cultivar, and more typical of other cultivars. The chromosome-scale assembly, version 1.1, contains 711 scaffolds covering 346.0 Mbp, with a contig N50 of 84.4 kbp, a scaffold N50 of 34.4 Mbp, and an evidence-based gene set of 29,408 loci. Version 1.1 has 10x the scaffold N50 and 4x the contig N50 as Criollo, and includes 111 Mb more anchored sequence. The version 1.1 assembly has 4.4% gap sequence, while Criollo has 10.9%. Through a combination of haplotype, association mapping and gene expression analyses, we leverage this robust reference genome to identify a promising candidate gene responsible for pod color variation. We demonstrate that green/red pod color in cacao is likely regulated by the R2R3 MYB transcription factor TcMYB113, homologs of which determine pigmentation in Rosaceae, Solanaceae, and Brassicaceae. One SNP within the target site for a highly conserved trans-acting siRNA in dicots, found within TcMYB113, seems to affect transcript levels of this gene and therefore pod color variation. Conclusions We report a high-quality sequence and annotation of Theobroma cacao L. and demonstrate its utility in identifying candidate genes regulating traits. PMID:23731509

Trichoderma theobromicola and T. paucisporum: two new species isolated from cacao in South America.

PubMed

Samuels, Gary J; Suarez, Carmen; Solis, Karina; Holmes, Keith A; Thomas, Sarah E; Ismaiel, Adnan; Evans, Harry C

2006-04-01

Trichoderma theobromicola and T. paucisporum spp. nov. are described. Trichoderma theobromicola was isolated as an endophyte from the trunk of a healthy cacao tree (Theobroma cacao, Malvaceae) in Amazonian Peru; it sporulates profusely on common mycological media. Trichoderma paucisporum is represented by two cultures that were obtained in Ecuador from cacao pods partially infected with frosty pod rot, Moniliophthora roreri; it sporulates sporadically and most cultures remain sterile on common media and autoclaved rice. It sporulates more reliably on synthetic low-nutrient agar (SNA) but produces few conidia. Trichoderma theobromicola was reintroduced into cacao seedlings through shoot inoculation and was recovered from stems but not from leaves, indicating that it is an endophytic species. Both produced a volatile/diffusable antibiotic that inhibited development of M. roreri in vitro and on-pod trials. Neither species demonstrated significant direct in vitro mycoparasitic activity against M. roreri.

Variability and correlation of physical attributes of soils cultivated with cacao trees in two climate zones in southern Bahia, Brazil

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao) is a very important crop in southern Bahia, Brazil, which needs good climate and soil conditions and management for great productivity. In this region, the culture is developed in a large variety of soils, which indicates differentiated products. The aim of this study was to ...

PCR-based identification of cacao black pod causal agents and identification of biological factors possibly contributing to Phytophthora megakarya's field dominance in West Africa

USDA-ARS?s Scientific Manuscript database

Among the Phytophthora species that cause black pod of cacao, P. megakarya is the most virulent, posing a serious threat to cacao production in Africa. Correct identification of the species causing the black pod and understanding the virulence factors involved are important for developing sustainabl...

Biological control of Black Pod Disease and Seedling Blight of cacao caused by Phytophthora Species using Trichoderma from Aceh Sumatra

USDA-ARS?s Scientific Manuscript database

The cocoa tree, Theobroma cacao L., suffers large yield losses in Aceh Indonesia to the disease black pod rot, caused by Phytophthora spp. Despite having the largest area under cacao production in Sumatra, farmers in the Aceh region have low overall production because of losses to insect pests and b...

Assessing genetic diversity in java fine-flavor cocoa (theobroma cacao l.) Germplasm by simple sequence repeat (ssr) markers

USDA-ARS?s Scientific Manuscript database

Indonesia is the 3rd largest cocoa producing countries in the world, with an annual cacao bean production of 572,000 tons. The currently cultivated cacao varieties in Indonesia were inter-hybrids of various clones introduced from the Americas since the 16th century. Among them, “Java cocoa” is a wel...

Nitrogen forms and levels influence on growth and nutrition of cacao

USDA-ARS?s Scientific Manuscript database

Ammonium and nitrate are the major forms of N present in tropical soils. A climatically controlled growth chamber experiment was conducted to assess the influence of forms (NO3-, NH4+, and mix of NO3- + NH4+) and levels (1.5 to 12.0 mM) of N on the growth and nutrition of cacao (Theobroma cacao L). ...

Genetic identity and diversity of Nigerian cacao genebank collections verified by Single Nucleotide polymorphisms (SNPs): A guide to field genebank management and utilization

USDA-ARS?s Scientific Manuscript database

Nigeria is the fourth largest cacao producer in the world. Field performance and quality of cacao hybrid families is largely dependent on the genetic integrity of parental clones obtained from field genebank collections. However, information on the impact of mislabeling on seed garden output and ger...

Population densities and genetic diversity of actinomycetes associated to the rhizosphere of Theobroma cacao

PubMed Central

Barreto, Tâmara R.; da Silva, Augusto C.M.; Soares, Ana Cristina F.; de Souza, Jorge T.

2008-01-01

In spite of the acknowledged importance of growth-promoting bacteria, only a reduced number of studies were conducted with these microorganisms on Theobroma cacao. The objectives of this work were to study the population densities and genetic diversity of actinomycetes associated with the rhizosphere of cacao as a first step in their application in plant growth promotion and biological control. The populations densities of actinomycetes in soil and cacao roots were similar, with mean values of 1,0 x 106 CFU/g and 9,6 x 105 CFU/g, respectively. All isolates selected and used in this study were identified through sequencing analyses of a fragment of the rpoB gene that encodes the β-subunit of the RNA polymerase as species of the genus Streptomyces. In vitro cellulolytic, xilanolytic and chitinolytic activity, indolacetic acid production and phosphate solubilization activities were observed in most of the isolates tested. The data obtained in this study demonstrate that actinomycetes account for a higher percentage of the total population of culturable bacteria in soil than on cacao roots. Additionally, actinomycetes from the cacao rhizosphere are genetically diverse and have potential applications as agents of growth promotion. PMID:24031247

A genetically anchored physical framework for Theobroma cacao cv. Matina 1-6

PubMed Central

2011-01-01

Background The fermented dried seeds of Theobroma cacao (cacao tree) are the main ingredient in chocolate. World cocoa production was estimated to be 3 million tons in 2010 with an annual estimated average growth rate of 2.2%. The cacao bean production industry is currently under threat from a rise in fungal diseases including black pod, frosty pod, and witches' broom. In order to address these issues, genome-sequencing efforts have been initiated recently to facilitate identification of genetic markers and genes that could be utilized to accelerate the release of robust T. cacao cultivars. However, problems inherent with assembly and resolution of distal regions of complex eukaryotic genomes, such as gaps, chimeric joins, and unresolvable repeat-induced compressions, have been unavoidably encountered with the sequencing strategies selected. Results Here, we describe the construction of a BAC-based integrated genetic-physical map of the T. cacao cultivar Matina 1-6 which is designed to augment and enhance these sequencing efforts. Three BAC libraries, each comprised of 10× coverage, were constructed and fingerprinted. 230 genetic markers from a high-resolution genetic recombination map and 96 Arabidopsis-derived conserved ortholog set (COS) II markers were anchored using pooled overgo hybridization. A dense tile path consisting of 29,383 BACs was selected and end-sequenced. The physical map consists of 154 contigs and 4,268 singletons. Forty-nine contigs are genetically anchored and ordered to chromosomes for a total span of 307.2 Mbp. The unanchored contigs (105) span 67.4 Mbp and therefore the estimated genome size of T. cacao is 374.6 Mbp. A comparative analysis with A. thaliana, V. vinifera, and P. trichocarpa suggests that comparisons of the genome assemblies of these distantly related species could provide insights into genome structure, evolutionary history, conservation of functional sites, and improvements in physical map assembly. A comparison between the two T. cacao cultivars Matina 1-6 and Criollo indicates a high degree of collinearity in their genomes, yet rearrangements were also observed. Conclusions The results presented in this study are a stand-alone resource for functional exploitation and enhancement of Theobroma cacao but are also expected to complement and augment ongoing genome-sequencing efforts. This resource will serve as a template for refinement of the T. cacao genome through gap-filling, targeted re-sequencing, and resolution of repetitive DNA arrays. PMID:21846342

A genetically anchored physical framework for Theobroma cacao cv. Matina 1-6.

PubMed

Saski, Christopher A; Feltus, Frank A; Staton, Margaret E; Blackmon, Barbara P; Ficklin, Stephen P; Kuhn, David N; Schnell, Raymond J; Shapiro, Howard; Motamayor, Juan Carlos

2011-08-16

The fermented dried seeds of Theobroma cacao (cacao tree) are the main ingredient in chocolate. World cocoa production was estimated to be 3 million tons in 2010 with an annual estimated average growth rate of 2.2%. The cacao bean production industry is currently under threat from a rise in fungal diseases including black pod, frosty pod, and witches' broom. In order to address these issues, genome-sequencing efforts have been initiated recently to facilitate identification of genetic markers and genes that could be utilized to accelerate the release of robust T. cacao cultivars. However, problems inherent with assembly and resolution of distal regions of complex eukaryotic genomes, such as gaps, chimeric joins, and unresolvable repeat-induced compressions, have been unavoidably encountered with the sequencing strategies selected. Here, we describe the construction of a BAC-based integrated genetic-physical map of the T. cacao cultivar Matina 1-6 which is designed to augment and enhance these sequencing efforts. Three BAC libraries, each comprised of 10× coverage, were constructed and fingerprinted. 230 genetic markers from a high-resolution genetic recombination map and 96 Arabidopsis-derived conserved ortholog set (COS) II markers were anchored using pooled overgo hybridization. A dense tile path consisting of 29,383 BACs was selected and end-sequenced. The physical map consists of 154 contigs and 4,268 singletons. Forty-nine contigs are genetically anchored and ordered to chromosomes for a total span of 307.2 Mbp. The unanchored contigs (105) span 67.4 Mbp and therefore the estimated genome size of T. cacao is 374.6 Mbp. A comparative analysis with A. thaliana, V. vinifera, and P. trichocarpa suggests that comparisons of the genome assemblies of these distantly related species could provide insights into genome structure, evolutionary history, conservation of functional sites, and improvements in physical map assembly. A comparison between the two T. cacao cultivars Matina 1-6 and Criollo indicates a high degree of collinearity in their genomes, yet rearrangements were also observed. The results presented in this study are a stand-alone resource for functional exploitation and enhancement of Theobroma cacao but are also expected to complement and augment ongoing genome-sequencing efforts. This resource will serve as a template for refinement of the T. cacao genome through gap-filling, targeted re-sequencing, and resolution of repetitive DNA arrays.

Repression of calcitonin gene-related peptide expression in trigeminal neurons by a Theobroma cacao extract☆

PubMed Central

Abbey, Marcie J.; Patil, Vinit V.; Vause, Carrie V.; Durham, Paul L.

2008-01-01

Ethnopharmacological relevance Cocoa bean preparations were first used by the ancient Maya and Aztec civilizations of South America to treat a variety of medical ailments involving the cardiovascular, gastrointestinal, and nervous systems. Diets rich in foods containing abundant polyphenols, as found in cocoa, underlie the protective effects reported in chronic inflammatory diseases. Release of calcitonin gene-related peptide (CGRP) from trigeminal nerves promotes inflammation in peripheral tissues and nociception. Aim of the study To determine whether a methanol extract of Theobroma cacao L. (Sterculiaceae) beans enriched for polyphenols could inhibit CGRP expression, both an in vitro and an in vivo approach was taken. Results Treatment of rat trigeminal ganglia cultures with depolarizing stimuli caused a significant increase in CGRP release that was repressed by pretreatment with Theobroma cacao extract. Pretreatment with Theobroma cacao was also shown to block the KCl- and capsaicin-stimulated increases in intracellular calcium. Next, the effects of Theobroma cacao on CGRP levels were determined using an in vivo model of temporomandibular joint (TMJ) inflammation. Capsaicin injection into the TMJ capsule caused an ipsilateral decrease in CGRP levels. Theobroma cacao extract injected into the TMJ capsule 24 h prior to capsaicin treatment repressed the stimulatory effects of capsaicin. Conclusions Our results demonstrate that Theobroma cacao extract can repress stimulated CGRP release by a mechanism that likely involves blockage of calcium channel activity. Furthermore, our findings suggest that the beneficial effects of diets rich in cocoa may include suppression of sensory trigeminal nerve activation. PMID:17997062

Theobroma cacao cystatins impair Moniliophthora perniciosa mycelial growth and are involved in postponing cell death symptoms.

PubMed

Pirovani, Carlos Priminho; da Silva Santiago, André; dos Santos, Lívia Santana; Micheli, Fabienne; Margis, Rogério; da Silva Gesteira, Abelmon; Alvim, Fátima Cerqueira; Pereira, Gonçalo Amarante Guimarães; de Mattos Cascardo, Júlio Cézar

2010-11-01

Three cystatin open reading frames named TcCys1, TcCys2 and TcCys3 were identified in cDNA libraries from compatible interactions between Theobroma cacao (cacao) and Moniliophthora perniciosa. In addition, an ORF named TcCys4 was identified in the cDNA library of the incompatible interaction. The cDNAs encoded conceptual proteins with 209, 127, 124, and 205 amino acid residues, with a deduced molecular weight of 24.3, 14.1, 14.3 and 22.8 kDa, respectively. His-tagged recombinant proteins were purified from Escherichia coli expression, and showed inhibitory activities against M. perniciosa. The four recombinant cystatins exhibited K(i) values against papain in the range of 152-221 nM. Recombinant TcCYS3 and TcCYS4 immobilized in CNBr-Sepharose were efficient to capture M. perniciosa proteases from culture media. Polyclonal antibodies raised against the recombinant TcCYS4 detected that the endogenous protein was more abundant in young cacao tissues, when compared with mature tissues. A ~85 kDa cacao multicystatin induced by M. perniciosa inoculation, MpNEP (necrosis and ethylene-inducing protein) and M. perniciosa culture supernatant infiltration were detected by anti-TcCYS4 antibodies in cacao young tissues. A direct role of the cacao cystatins in the defense against this phytopathogen was proposed, as well as its involvement in the development of symptoms of programmed cell death.

Repression of calcitonin gene-related peptide expression in trigeminal neurons by a Theobroma cacao extract.

PubMed

Abbey, Marcie J; Patil, Vinit V; Vause, Carrie V; Durham, Paul L

2008-01-17

Cocoa bean preparations were first used by the ancient Maya and Aztec civilizations of South America to treat a variety of medical ailments involving the cardiovascular, gastrointestinal, and nervous systems. Diets rich in foods containing abundant polyphenols, as found in cocoa, underlie the protective effects reported in chronic inflammatory diseases. Release of calcitonin gene-related peptide (CGRP) from trigeminal nerves promotes inflammation in peripheral tissues and nociception. To determine whether a methanol extract of Theobroma cacao L. (Sterculiaceae) beans enriched for polyphenols could inhibit CGRP expression, both an in vitro and an in vivo approach was taken. Treatment of rat trigeminal ganglia cultures with depolarizing stimuli caused a significant increase in CGRP release that was repressed by pretreatment with Theobroma cacao extract. Pretreatment with Theobroma cacao was also shown to block the KCl- and capsaicin-stimulated increases in intracellular calcium. Next, the effects of Theobroma cacao on CGRP levels were determined using an in vivo model of temporomandibular joint (TMJ) inflammation. Capsaicin injection into the TMJ capsule caused an ipsilateral decrease in CGRP levels. Theobroma cacao extract injected into the TMJ capsule 24h prior to capsaicin treatment repressed the stimulatory effects of capsaicin. Our results demonstrate that Theobroma cacao extract can repress stimulated CGRP release by a mechanism that likely involves blockage of calcium channel activity. Furthermore, our findings suggest that the beneficial effects of diets rich in cocoa may include suppression of sensory trigeminal nerve activation.

Transposon fingerprinting using low coverage whole genome shotgun sequencing in cacao (Theobroma cacao L.) and related species.

PubMed

Sveinsson, Saemundur; Gill, Navdeep; Kane, Nolan C; Cronk, Quentin

2013-07-24

Transposable elements (TEs) and other repetitive elements are a large and dynamically evolving part of eukaryotic genomes, especially in plants where they can account for a significant proportion of genome size. Their dynamic nature gives them the potential for use in identifying and characterizing crop germplasm. However, their repetitive nature makes them challenging to study using conventional methods of molecular biology. Next generation sequencing and new computational tools have greatly facilitated the investigation of TE variation within species and among closely related species. (i) We generated low-coverage Illumina whole genome shotgun sequencing reads for multiple individuals of cacao (Theobroma cacao) and related species. These reads were analysed using both an alignment/mapping approach and a de novo (graph based clustering) approach. (ii) A standard set of ultra-conserved orthologous sequences (UCOS) standardized TE data between samples and provided phylogenetic information on the relatedness of samples. (iii) The mapping approach proved highly effective within the reference species but underestimated TE abundance in interspecific comparisons relative to the de novo methods. (iv) Individual T. cacao accessions have unique patterns of TE abundance indicating that the TE composition of the genome is evolving actively within this species. (v) LTR/Gypsy elements are the most abundant, comprising c.10% of the genome. (vi) Within T. cacao the retroelement families show an order of magnitude greater sequence variability than the DNA transposon families. (vii) Theobroma grandiflorum has a similar TE composition to T. cacao, but the related genus Herrania is rather different, with LTRs making up a lower proportion of the genome, perhaps because of a massive presence (c. 20%) of distinctive low complexity satellite-like repeats in this genome. (i) Short read alignment/mapping to reference TE contigs provides a simple and effective method of investigating intraspecific differences in TE composition. It is not appropriate for comparing repetitive elements across the species boundaries, for which de novo methods are more appropriate. (ii) Individual T. cacao accessions have unique spectra of TE composition indicating active evolution of TE abundance within this species. TE patterns could potentially be used as a "fingerprint" to identify and characterize cacao accessions.

Carbon and water fluxes above a cacao plantation in Sulawesi, Indonesia

NASA Astrophysics Data System (ADS)

Falk, U.; Ibrom, A.

2003-04-01

The investigation of interactions between biosphere and atmosphere of the major land use types of the tropical rain forest margin area in South East Asia and quantification of the impact that land use change from undisturbed primary rain forest to pasture has on these interactions is task of subprogramme B1 within the DFG-funded project STORMA (Stability of Rain Forest Margins). In order to fulfill the projects tasks the different major land use types have to be investigated and each ecosystem characterized one by one and compared to a reference site in an undisturbed primary rain forest, to see the changes in the atmosphere-biospheric interactions, i. e. in water and carbon household, with land use change and thus the impact on regional climate. One of the major land use types in the valleys around the Lore Lindu National Park on Sulawesi are Cacao plantations, Theobroma cacao. A site in the Palolo valley near the village Nopu was chosen as research site since the area there is covered with small Cacao fields which form to one big area of Cacao and matches the requirements of the applied research approach. Since Cacao trees need to be shaded especially when younger, shadow trees had been planted and trees of the former forest had been left standing to serve as wind breaks and sun shades. The plantations in Nopu, Palolo valley, consist not only of fields of cultivated Cacao, but also serve as environment and home to the farmers and their families. The whole area of Cacao plantation is interspersed with wooden farm houses, which are also sources of carbon dioxide due to cooking or small power plants etc. and thus have to be taken into account when looking at the carbon household of this specific ecosystem. An estimation of the components of the carbon and water household and the contribution of the humans living within this environment to the carbon household of Cacao plantations of this ecosystem is subject of this presentation. From December 2001 until April 2002 and June 2002 until now eddy-covariance measurements have been performed above a Cacao plantation in Nopu measuring time series of water vapour, CO2, air temperature, three-dimensional wind vector, photosyntetic active radiation and the surface temperature of the Cacao canopy at 10 Hz. Additionally, net radiation balance and soil heat fluxes have been measured. In order to assess the carbon input caused by the humans living in the ecosystem, a mapping of the site area has been carried out, including investigations of consumption of fire wood and use of machines, like generators for example. In order to obtain the energy balance equation of the canopy surface, also the radiation balance and the heat flux into the canopy have to be evaluated.

Theobroma cacao extract attenuates the development of Dermatophagoides farinae-induced atopic dermatitis-like symptoms in NC/Nga mice.

PubMed

Kang, Heerim; Lee, Chang Hyung; Kim, Jong Rhan; Kwon, Jung Yeon; Son, Myoung-Jin; Kim, Jong-Eun; Lee, Ki Won

2017-02-01

Cacao beans from Theobroma cacao are an abundant source of polyphenols, particularly flavonoids. Previous studies demonstrated that cacao flavanols decrease pro-inflammatory cytokines resulting in the alleviation of allergic symptoms. We sought to investigate the effects of cacao extract (CE) on Dermatophagoides farinae extract (DFE)-induced atopic dermatitis (AD)-like symptoms. CE attenuated DFE-induced AD-like symptoms as assessed by skin lesion analyses, dermatitis score, and skin thickness. Histopathological analysis revealed that CE suppressed DFE-induced immune cell infiltration into the skin. These observations occurred concomitantly with the downregulation of inflammatory markers including serum immunoglobulin (Ig) E, chemokine; thymus and activation-regulated chemokine and macrophage-derived chemokine as well as the skin-derived cytokines interleukin (IL)-4, IL-5, and interferon-γ. CE also significantly alleviated transepidermal water loss and increased skin hydration. These results suggest that CE, a natural phytochemical-rich food, has potential therapeutic efficacy for the treatment of AD. Copyright © 2016. Published by Elsevier Ltd.

Identification and mapping of conserved ortholog set(COS) II sequences of cacao and their conversion to SNP markers for marker-assisted selection in Theobroma cocoa and comparative genomics studies

USDA-ARS?s Scientific Manuscript database

Theobroma cacao is a tree cultivated in the tropics around the world for its seeds that are the source of both chocolate and cocoa butter. The cacao genome sequencing project initiated as a collaboration between USDA, Mars, Inc. and IBM has generated a great deal of transcriptome and genome sequenc...

Combination of RNAseq and SNP nanofluidic array reveals the center of genetic diversity of cacao pathogen Moniliophthora roreri in the upper Magdalena Valley of Colombia and its clonality

USDA-ARS?s Scientific Manuscript database

Moniliophthora roreri is the fungal pathogen that causes frosty pod rot (FPR) disease of Theobroma cacao L., the source of chocolate. FPR occurs in most of the cacao producing countries in the Western Hemisphere, causing yield losses up to 80%. Genetic diversity within the FPR pathogen population ma...

Path analysis of phenotypic traits in young cacao plants under drought conditions.

PubMed

Santos, Emerson Alves Dos; Almeida, Alex-Alan Furtado de; Branco, Marcia Christina da Silva; Santos, Ivanildes Conceição Dos; Ahnert, Dario; Baligar, Virupax C; Valle, Raúl René

2018-01-01

Drought is worldwide considered one of the most limiting factors of Theobroma cacao production, which can be intensified by global climate changes. In this study, we aimed to investigate the phenotypic correlation among morphological characteristics of cacao progenies submitted to irrigation and drought conditions and their partitions into direct and indirect effects. Path analysis with phenotypic plasticity index was used as criteria for estimation of basic and explanatory variables. The experiment was conducted in a greenhouse at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a randomized block 21 x 2 factorial arrangement [21 cacao progenies obtained from complete diallel crosses and two water regimes (control and drought)] and six replications. In general, drought conditions influenced biomass production in most progenies, causing significant reductions in total leaf area, leaf number, leaf biomass, fine-roots length (diameter <1 mm), root volume and root area for considered drought intolerant. All progenies showed alterations in growth due to drought. Phenotypic plasticity was most strongly pronounced in root volume. Stem and root diameters, as well as stem dry biomass were the growth variables with the greatest direct effects on root volume under drought conditions, these characters being indicated in screening of cacao progenies drought tolerant.

Path analysis of phenotypic traits in young cacao plants under drought conditions

PubMed Central

dos Santos, Emerson Alves; de Almeida, Alex-Alan Furtado; Branco, Marcia Christina da Silva; dos Santos, Ivanildes Conceição; Ahnert, Dario; Baligar, Virupax C.; Valle, Raúl René

2018-01-01

Drought is worldwide considered one of the most limiting factors of Theobroma cacao production, which can be intensified by global climate changes. In this study, we aimed to investigate the phenotypic correlation among morphological characteristics of cacao progenies submitted to irrigation and drought conditions and their partitions into direct and indirect effects. Path analysis with phenotypic plasticity index was used as criteria for estimation of basic and explanatory variables. The experiment was conducted in a greenhouse at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a randomized block 21 x 2 factorial arrangement [21 cacao progenies obtained from complete diallel crosses and two water regimes (control and drought)] and six replications. In general, drought conditions influenced biomass production in most progenies, causing significant reductions in total leaf area, leaf number, leaf biomass, fine-roots length (diameter <1 mm), root volume and root area for considered drought intolerant. All progenies showed alterations in growth due to drought. Phenotypic plasticity was most strongly pronounced in root volume. Stem and root diameters, as well as stem dry biomass were the growth variables with the greatest direct effects on root volume under drought conditions, these characters being indicated in screening of cacao progenies drought tolerant. PMID:29408854

Infection of non-host model plant species with the narrow-host-range Cacao swollen shoot virus.

PubMed

Friscina, Arianna; Chiappetta, Laura; Jacquemond, Mireille; Tepfer, Mark

2017-02-01

Cacao swollen shoot virus (CSSV) is a major pathogen of cacao (Theobroma cacao) in Africa, and long-standing efforts to limit its spread by the culling of infected trees have had very limited success. CSSV is a particularly difficult virus to study, as it has a very narrow host range, limited to several tropical tree species. Furthermore, the virus is not mechanically transmissible, and its insect vector can only be used with difficulty. Thus, the only efficient means to infect cacao plants that have been experimentally described so far are by particle bombardment or the agroinoculation of cacao plants with an infectious clone. We have genetically transformed three non-host species with an infectious form of the CSSV genome: two experimental hosts widely used in plant virology (Nicotiana tabacum and N. benthamiana) and the model species Arabidopsis thaliana. In transformed plants of all three species, the CSSV genome was able to replicate, and, in tobacco, CSSV particles could be observed by immunosorbent electron microscopy, demonstrating that the complete virus cycle could be completed in a non-host plant. These results will greatly facilitate the preliminary testing of CSSV control strategies using plants that are easy to raise and to transform genetically. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Exposure to the leaf litter microbiome of healthy adults protects seedlings from pathogen damage.

PubMed

Christian, Natalie; Herre, Edward Allen; Mejia, Luis C; Clay, Keith

2017-07-12

It is increasingly recognized that microbiota affect host health and physiology. However, it is unclear what factors shape microbiome community assembly in nature, and how microbiome assembly can be manipulated to improve host health. All plant leaves host foliar endophytic fungi, which make up a diverse, environmentally acquired fungal microbiota. Here, we experimentally manipulated assembly of the cacao tree ( Theobroma cacao ) fungal microbiome in nature and tested the effect of assembly outcome on host health. Using next-generation sequencing, as well as culture-based methods coupled with Sanger sequencing, we found that manipulating leaf litter exposure and location within the forest canopy significantly altered microbiome composition in cacao. Exposing cacao seedlings to leaf litter from healthy conspecific adults enriched the seedling microbiome with Colletotrichum tropicale , a fungal endophyte known to enhance pathogen resistance of cacao seedlings by upregulating host defensive pathways. As a result, seedlings exposed to healthy conspecific litter experienced reduced pathogen damage. Our results link processes that affect the assembly and composition of microbiome communities to their functional consequences for host success, and have broad implications for understanding plant-microbe interactions. Deliberate manipulation of the plant-fungal microbiome also has potentially important applications for cacao production and other agricultural systems in general. © 2017 The Author(s).

Cacao diseases: a global perspective from an industry point of view.

PubMed

Hebbar, Prakash K

2007-12-01

ABSTRACT Diseases of cacao, Theobroma cacao, account for losses of more than 30% of the potential crop. These losses have caused a steady decline in production and a reduction in bean quality in almost all the cacao-producing areas in the world, especially in small-holder farms in Latin America and West Africa. The most significant diseases are witches' broom, caused by Moniliophthora perniciosa, which occurs mainly in South America; frosty pod rot, caused by M. roreri, which occurs mainly in Central and northern South America; and black pod disease, caused by several species of Phytophthora, which are distributed throughout the tropics. In view of the threat that these diseases pose to the sustainability of the cacao crop, Mars Inc. and their industry partners have funded collaborative research involving cacao research institutes and governmental and nongovernmental agencies. The objective of this global initiative is to develop short- to medium-term, low-cost, environmentally friendly disease-management strategies until disease tolerant varieties are widely available. These include good farming practices, biological control and the rational or minimal use of chemicals that could be used for integrated pest management (IPM). Farmer field schools are used to get these technologies to growers. This paper describes some of the key collaborative partners and projects that are underway in South America and West Africa.

Genetic diversity and parentage in farmer selections of cacao from Southern Sulawesi, Indonesia revealed by microsatellite markers.

PubMed

Dinarti, Diny; Susilo, Agung W; Meinhardt, Lyndel W; Ji, Kun; Motilal, Lambert A; Mischke, Sue; Zhang, Dapeng

2015-12-01

Indonesia is the third largest cocoa-producing country in the world. Knowledge of genetic diversity and parentage of farmer selections is important for effective selection and rational deployment of superior cacao clones in farmers' fields. We assessed genetic diversity and parentage of 53 farmer selections of cacao in Sulawesi, Indonesia, using 152 international clones as references. Cluster analysis, based on 15 microsatellite markers, showed that these Sulawesi farmer selections are mainly comprised of hybrids derived from Trinitario and two Upper Amazon Forastero groups. Bayesian assignment and likelihood-based parentage analysis further demonstrated that only a small number of germplasm groups, dominantly Trinitario and Parinari, contributed to these farmer selections, in spite of diverse parental clones having been used in the breeding program and seed gardens in Indonesia since the 1950s. The narrow parentage predicts a less durable host resistance to cacao diseases. Limited access of the farmers to diverse planting materials or the strong preference for large pods and large bean size by local farmers, may have affected the selection outcome. Diverse sources of resistance, harbored in different cacao germplasm groups, need to be effectively incorporated to broaden the on-farm diversity and ensure sustainable cacao production in Sulawesi.

Sulawesi cacao (Theobroma cacao, l.) performances under two different agricultural system in east coast of Central Sulawesi

NASA Astrophysics Data System (ADS)

Muslimin; Wijayanti, W.; Anshary, A.; Basri, Z.; Cruz, A. F.; Suwastika, I. N.; Shiina, T.

2018-04-01

Cacao (Theobroma cacao L.) is one major agricultural commodity from central Sulawesi, which is significantly affected by a range of pest and diseases including Cacao Pod Borer (CPB), Pod Rod Diseases (PRD), and Vascular Streak Dieback (VSD). Intensive and extensive approaches, including introduction of various new superior cultivars/clones, and development of eco-friendly agricultural system were implemented in order to overtake that problem. Here, we observed the performance of three different clones (namely: Sulawesi 1, MCC01, and Tadulako-1) in two different locations (Parigi and Poso-which located along east coast of central Sulawesi province), and under two different Agricultural management system (echo-friendly-intensive and Non-Intensive). All clones tested showed their well-adapted and suitable to local conditions. The performance of each clone can be improved by intensive management system. Based on all observed parameters (number of wet bean per pot, bean count, fat and shell percentage, and percentage of infection) it convincingly showed that intensive management system was working well in improving the quality and quantity of cacao beans production, and it fit to commercial requirements. The highest rate of infection was by Phytophthora, and no significant differences on the overall performances. Clones of Sulawesi-1 and MCC01 most likely was better than Tadulako-1.

Chromatin differentiation between Theobroma cacao L. and T. grandiflorum Schum

PubMed Central

2010-01-01

A comparative analysis of mitotic chromosomes of Theobroma cacao (cacao) and T. grandiflorum (cupuaçu) was performed aiming to identify cytological differences between the two most important species of this genus. Both species have symmetric karyotypes, with 2n = 20 metacentric chromosomes ranging in size from 2.00 to 1.19 μm (cacao) and from 2.21 to 1.15 μm (cupuaçu). The interphase nuclei of both species were of the arreticulate type, displaying up to 20 chromocentres, which were more regularly shaped in cacao than in cupuaçu. Prophase chromosomes of both species were more condensed in the proximal region, sometimes including the whole short arm. Both species exhibited only one pair of terminal heterochromatic bands, positively stained with chromomycin A 3 , which co-localized with the single 45S rDNA site. Each karyotype displayed a single 5S rDNA site in the proximal region of another chromosome pair. Heterochromatic bands were also observed on the centromeric/pericentromeric regions of all 20 chromosomes of cacao after C-banding followed by Giemsa or DAPI staining, whereas in cupuaçu they were never detected. These data suggest that the chromosomes of both species have been largely conserved and their pericentromeric chromatin is the only citologically differentiated region. PMID:21637611

Fermentation of cacao (Theobroma cacao L.) seeds with a hybrid Kluyveromyces marxianus strain improved product quality attributes.

PubMed

Leal, Gildemberg Amorim; Gomes, Luiz Humberto; Efraim, Priscilla; de Almeida Tavares, Flavio Cesar; Figueira, Antonio

2008-08-01

Fermentation of Theobroma cacao (cacao) seeds is an absolute requirement for the full development of chocolate flavor precursors. An adequate aeration of the fermenting cacao seed mass is a fundamental prerequisite for a satisfactory fermentation. Here, we evaluated whether a controlled inoculation of cacao seed fermentation using a Kluyveromyces marxianus hybrid yeast strain, with an increased pectinolytic activity, would improve an earlier liquid drainage ('sweatings') from the fermentation mass, developing a superior final product quality. Inoculation with K. marxianus increased by one third the volume of drained liquid and affected the microorganism population structure during fermentation, which was detectable up to the end of the process. Introduction of the hybrid yeast affected the profile of total seed protein degradation evaluated by polyacrylamide gel electrophoresis, with improved seed protein degradation, and reduction of titrable acidity. Sensorial evaluation of the chocolate obtained from beans fermented with the K. marxianus inoculation was more accepted by analysts in comparison with the one from cocoa obtained through natural fermentation. The increase in mass aeration during the first 24 h seemed to be fundamental for the improvement of fermentation quality, demonstrating the potential application of this improved hybrid yeast strain with superior exogenous pectinolytic activity.

Genetic diversity and parentage in farmer selections of cacao from Southern Sulawesi, Indonesia revealed by microsatellite markers

PubMed Central

Dinarti, Diny; Susilo, Agung W.; Meinhardt, Lyndel W.; Ji, Kun; Motilal, Lambert A.; Mischke, Sue; Zhang, Dapeng

2015-01-01

Indonesia is the third largest cocoa-producing country in the world. Knowledge of genetic diversity and parentage of farmer selections is important for effective selection and rational deployment of superior cacao clones in farmers’ fields. We assessed genetic diversity and parentage of 53 farmer selections of cacao in Sulawesi, Indonesia, using 152 international clones as references. Cluster analysis, based on 15 microsatellite markers, showed that these Sulawesi farmer selections are mainly comprised of hybrids derived from Trinitario and two Upper Amazon Forastero groups. Bayesian assignment and likelihood-based parentage analysis further demonstrated that only a small number of germplasm groups, dominantly Trinitario and Parinari, contributed to these farmer selections, in spite of diverse parental clones having been used in the breeding program and seed gardens in Indonesia since the 1950s. The narrow parentage predicts a less durable host resistance to cacao diseases. Limited access of the farmers to diverse planting materials or the strong preference for large pods and large bean size by local farmers, may have affected the selection outcome. Diverse sources of resistance, harbored in different cacao germplasm groups, need to be effectively incorporated to broaden the on-farm diversity and ensure sustainable cacao production in Sulawesi. PMID:26719747

Genome-wide identification and characterization of cacao WRKY transcription factors and analysis of their expression in response to witches' broom disease

PubMed Central

Silva Monteiro de Almeida, Dayanne; Oliveira Jordão do Amaral, Daniel; Del-Bem, Luiz-Eduardo; Bronze dos Santos, Emily; Santana Silva, Raner José; Peres Gramacho, Karina; Vincentz, Michel

2017-01-01

Transcriptional regulation, led by transcription factors (TFs) such as those of the WRKY family, is a mechanism used by the organism to enhance or repress gene expression in response to stimuli. Here, we report on the genome-wide analysis of the Theobroma cacao WRKY TF family and also investigate the expression of WRKY genes in cacao infected by the fungus Moniliophthora perniciosa. In the cacao genome, 61 non-redundant WRKY sequences were found and classified in three groups (I to III) according to the WRKY and zinc-finger motif types. The 61 putative WRKY sequences were distributed on the 10 cacao chromosomes and 24 of them came from duplication events. The sequences were phylogenetically organized according to the general WRKY groups. The phylogenetic analysis revealed that subgroups IIa and IIb are sister groups and share a common ancestor, as well as subgroups IId and IIe. The most divergent groups according to the plant origin were IIc and III. According to the phylogenetic analysis, 7 TcWRKY genes were selected and analyzed by RT-qPCR in susceptible and resistant cacao plants infected (or not) with M. perniciosa. Some TcWRKY genes presented interesting responses to M. perniciosa such as Tc01_p014750/Tc06_p013130/AtWRKY28, Tc09_p001530/Tc06_p004420/AtWRKY40, Tc04_p016130/AtWRKY54 and Tc10_p016570/ AtWRKY70. Our results can help to select appropriate candidate genes for further characterization in cacao or in other Theobroma species. PMID:29084273

Carbon Storage in Soil Size Fractions Under Two Cacao Agroforestry Systems in Bahia, Brazil

NASA Astrophysics Data System (ADS)

Gama-Rodrigues, Emanuela F.; Ramachandran Nair, P. K.; Nair, Vimala D.; Gama-Rodrigues, Antonio C.; Baligar, Virupax C.; Machado, Regina C. R.

2010-02-01

Shaded perennial agroforestry systems contain relatively high quantities of soil carbon (C) resulting from continuous deposition of plant residues; however, the extent to which the C is sequestered in soil will depend on the extent of physical protection of soil organic C (SOC). The main objective of this study was to characterize SOC storage in relation to soil fraction-size classes in cacao ( Theobroma cacao L.) agroforestry systems (AFSs). Two shaded cacao systems and an adjacent natural forest in reddish-yellow Oxisols in Bahia, Brazil were selected. Soil samples were collected from four depth classes to 1 m depth and separated by wet-sieving into three fraction-size classes (>250 μm, 250-53 μm, and <53 μm)—corresponding to macroaggregate, microaggregate, and silt-and-clay size fractions—and analyzed for C content. The total SOC stock did not vary among systems (mean: 302 Mg/ha). On average, 72% of SOC was in macroaggregate-size, 20% in microaggregate-size, and 8% in silt-and-clay size fractions in soil. Sonication of aggregates showed that occlusion of C in soil aggregates could be a major mechanism of C protection in these soils. Considering the low level of soil disturbances in cacao AFSs, the C contained in the macroaggregate fraction might become stabilized in the soil. The study shows the role of cacao AFSs in mitigating greenhouse gas (GHG) emission through accumulation and retention of high amounts of organic C in the soils and suggests the potential benefit of this environmental service to the nearly 6 million cacao farmers worldwide.

Genome-wide identification and characterization of cacao WRKY transcription factors and analysis of their expression in response to witches' broom disease.

PubMed

Silva Monteiro de Almeida, Dayanne; Oliveira Jordão do Amaral, Daniel; Del-Bem, Luiz-Eduardo; Bronze Dos Santos, Emily; Santana Silva, Raner José; Peres Gramacho, Karina; Vincentz, Michel; Micheli, Fabienne

2017-01-01

Transcriptional regulation, led by transcription factors (TFs) such as those of the WRKY family, is a mechanism used by the organism to enhance or repress gene expression in response to stimuli. Here, we report on the genome-wide analysis of the Theobroma cacao WRKY TF family and also investigate the expression of WRKY genes in cacao infected by the fungus Moniliophthora perniciosa. In the cacao genome, 61 non-redundant WRKY sequences were found and classified in three groups (I to III) according to the WRKY and zinc-finger motif types. The 61 putative WRKY sequences were distributed on the 10 cacao chromosomes and 24 of them came from duplication events. The sequences were phylogenetically organized according to the general WRKY groups. The phylogenetic analysis revealed that subgroups IIa and IIb are sister groups and share a common ancestor, as well as subgroups IId and IIe. The most divergent groups according to the plant origin were IIc and III. According to the phylogenetic analysis, 7 TcWRKY genes were selected and analyzed by RT-qPCR in susceptible and resistant cacao plants infected (or not) with M. perniciosa. Some TcWRKY genes presented interesting responses to M. perniciosa such as Tc01_p014750/Tc06_p013130/AtWRKY28, Tc09_p001530/Tc06_p004420/AtWRKY40, Tc04_p016130/AtWRKY54 and Tc10_p016570/ AtWRKY70. Our results can help to select appropriate candidate genes for further characterization in cacao or in other Theobroma species.

Carbon storage in soil size fractions under two cacao agroforestry systems in Bahia, Brazil.

PubMed

Gama-Rodrigues, Emanuela F; Ramachandran Nair, P K; Nair, Vimala D; Gama-Rodrigues, Antonio C; Baligar, Virupax C; Machado, Regina C R

2010-02-01

Shaded perennial agroforestry systems contain relatively high quantities of soil carbon (C) resulting from continuous deposition of plant residues; however, the extent to which the C is sequestered in soil will depend on the extent of physical protection of soil organic C (SOC). The main objective of this study was to characterize SOC storage in relation to soil fraction-size classes in cacao (Theobroma cacao L.) agroforestry systems (AFSs). Two shaded cacao systems and an adjacent natural forest in reddish-yellow Oxisols in Bahia, Brazil were selected. Soil samples were collected from four depth classes to 1 m depth and separated by wet-sieving into three fraction-size classes (>250 microm, 250-53 microm, and <53 microm)-corresponding to macroaggregate, microaggregate, and silt-and-clay size fractions-and analyzed for C content. The total SOC stock did not vary among systems (mean: 302 Mg/ha). On average, 72% of SOC was in macroaggregate-size, 20% in microaggregate-size, and 8% in silt-and-clay size fractions in soil. Sonication of aggregates showed that occlusion of C in soil aggregates could be a major mechanism of C protection in these soils. Considering the low level of soil disturbances in cacao AFSs, the C contained in the macroaggregate fraction might become stabilized in the soil. The study shows the role of cacao AFSs in mitigating greenhouse gas (GHG) emission through accumulation and retention of high amounts of organic C in the soils and suggests the potential benefit of this environmental service to the nearly 6 million cacao farmers worldwide.

Application of Genome Wide Association and Genomic Prediction for Improvement of Cacao Productivity and Resistance to Black and Frosty Pod Diseases

PubMed Central

Romero Navarro, J. Alberto; Phillips-Mora, Wilbert; Arciniegas-Leal, Adriana; Mata-Quirós, Allan; Haiminen, Niina; Mustiga, Guiliana; Livingstone III, Donald; van Bakel, Harm; Kuhn, David N.; Parida, Laxmi; Kasarskis, Andrew; Motamayor, Juan C.

2017-01-01

Chocolate is a highly valued and palatable confectionery product. Chocolate is primarily made from the processed seeds of the tree species Theobroma cacao. Cacao cultivation is highly relevant for small-holder farmers throughout the tropics, yet its productivity remains limited by low yields and widespread pathogens. A panel of 148 improved cacao clones was assembled based on productivity and disease resistance, and phenotypic single-tree replicated clonal evaluation was performed for 8 years. Using high-density markers, the diversity of clones was expressed relative to 10 known ancestral cacao populations, and significant effects of ancestry were observed in productivity and disease resistance. Genome-wide association (GWA) was performed, and six markers were significantly associated with frosty pod disease resistance. In addition, genomic selection was performed, and consistent with the observed extensive linkage disequilibrium, high predictive ability was observed at low marker densities for all traits. Finally, quantitative trait locus mapping and differential expression analysis of two cultivars with contrasting disease phenotypes were performed to identify genes underlying frosty pod disease resistance, identifying a significant quantitative trait locus and 35 differentially expressed genes using two independent differential expression analyses. These results indicate that in breeding populations of heterozygous and recently admixed individuals, mapping approaches can be used for low complexity traits like pod color cacao, or in other species single gene disease resistance, however genomic selection for quantitative traits remains highly effective relative to mapping. Our results can help guide the breeding process for sustainable improved cacao productivity. PMID:29184558

Colletotrichum gloeosporioides s.l. associated with Theobroma cacao and other plants in Panama: multilocus phylogenies distinguish host-associated pathogens from asymptomatic endophytes.

PubMed

Rojas, Enith I; Rehner, Stephen A; Samuels, Gary J; Van Bael, Sunshine A; Herre, Edward A; Cannon, Paul; Chen, Rui; Pang, Junfeng; Wang, Ruiwu; Zhang, Yaping; Peng, Yan-Qiong; Sha, Tao

2010-01-01

Colletotrichum interacts with numerous plant species overtly as symptomatic pathogens and cryptically as asymptomatic endophytes. It is not known whether these contrasting ecological modes are optional strategies expressed by individual Colletotrichum species or whether a species' ecology is explicitly pathogenic or endophytic. We explored this question by inferring relationships among 77 C. gloeosporioides s.l. strains isolated from asymptomatic leaves and from anthracnose lesions on leaves and fruits of Theobroma cacao (cacao) and other plants from Panamá. ITS and 5'-tef1 were used to assess diversity and to delineate operational taxonomic units for multilocus phylogenetic analysis. The ITS and 5'-tef1 screens concordantly resolved four strongly supported lineages, clades A-D: Clade A includes the ex type of C. gloeosporioides, clade B includes the ex type ITS sequence of C. boninense, and clades C and D are unidentified. The ITS yielded limited resolution and support within all clades, in particular the C. gloeosporioides clade (A), the focal lineage dealt with in this study. In contrast the 5'-tef1 screen differentiated nine distinctive haplotype subgroups within the C. gloeosporioides clade that were concordant with phylogenetic terminals resolved in a five-locus nuclear phylogeny. Among these were two phylogenetic species associated with symptomatic infections specific to either cacao or mango and five phylogenetic species isolated principally as asymptomatic infections from cacao and other plant hosts. We formally describe two new species, C. tropicale and C. ignotum, that are frequent asymptomatic associates of cacao and other Neotropical plant species, and epitypify C. theobromicola, which is associated with foliar and fruit anthracnose lesions of cacao. Asymptomatic Colletotrichum strains isolated from cacao plants grown in China included six distinct C. gloeosporioides clade taxa, only one of which is known to occur in the Neotropics.

Changes in Soil Physical and Chemical Properties in Long Term Improved Natural and Traditional Agroforestry Management Systems of Cacao Genotypes in Peruvian Amazon

PubMed Central

Arévalo-Gardini, Enrique; Canto, Manuel; Alegre, Julio; Loli, Oscar; Julca, Alberto; Baligar, Virupax

2015-01-01

Growing cacao (Theobroma cacao L.) in an agroforestry system generates a productive use of the land, preserves the best conditions for physical, chemical and biological properties of tropical soils, and plays an important role in improving cacao production and fertility of degraded tropical soils. The aim of this study was to evaluate the impact of two long term agroforestry systems of cacao management on soil physical and chemical properties in an area originally inhabited by 30 years old native secondary forest (SF). The two agroforestry systems adapted were: improved natural agroforestry system (INAS) where trees without economic value were selectively removed to provide 50% shade and improved traditional agroforestry system (ITAS) where all native trees were cut and burnt in the location. For evaluation of the changes of soil physical and chemical properties with time due to the imposed cacao management systems, plots of 10 cacao genotypes (ICS95, UF613, CCN51, ICT1112, ICT1026, ICT2162, ICT2171, ICT2142, H35, U30) and one plot with a spontaneous hybrid were selected. Soil samples were taken at 0-20, 20-40 and 40-60 cm depths before the installation of the management systems (2004), and then followed at two years intervals. Bulk density, porosity, field capacity and wilting point varied significantly during the years of assessment in the different soil depths and under the systems assessed. Soil pH, CEC, exchangeable Mg and sum of the bases were higher in the INAS than the ITAS. In both systems, SOM, Ext. P, K and Fe, exch. K, Mg and Al+H decreased with years of cultivation; these changes were more evident in the 0-20 cm soil depth. Overall improvement of SOM and soil nutrient status was much higher in the ITAS than INAS. The levels of physical and chemical properties of soil under cacao genotypes showed a marked difference in both systems. PMID:26181053

Changes in soil physical and chemical properties in long term improved natural and traditional agroforestry management systems of cacao genotypes in Peruvian Amazon.

PubMed

Arévalo-Gardini, Enrique; Canto, Manuel; Alegre, Julio; Loli, Oscar; Julca, Alberto; Baligar, Virupax

2015-01-01

Growing cacao (Theobroma cacao L.) in an agroforestry system generates a productive use of the land, preserves the best conditions for physical, chemical and biological properties of tropical soils, and plays an important role in improving cacao production and fertility of degraded tropical soils. The aim of this study was to evaluate the impact of two long term agroforestry systems of cacao management on soil physical and chemical properties in an area originally inhabited by 30 years old native secondary forest (SF). The two agroforestry systems adapted were: improved natural agroforestry system (INAS) where trees without economic value were selectively removed to provide 50% shade and improved traditional agroforestry system (ITAS) where all native trees were cut and burnt in the location. For evaluation of the changes of soil physical and chemical properties with time due to the imposed cacao management systems, plots of 10 cacao genotypes (ICS95, UF613, CCN51, ICT1112, ICT1026, ICT2162, ICT2171, ICT2142, H35, U30) and one plot with a spontaneous hybrid were selected. Soil samples were taken at 0-20, 20-40 and 40-60 cm depths before the installation of the management systems (2004), and then followed at two years intervals. Bulk density, porosity, field capacity and wilting point varied significantly during the years of assessment in the different soil depths and under the systems assessed. Soil pH, CEC, exchangeable Mg and sum of the bases were higher in the INAS than the ITAS. In both systems, SOM, Ext. P, K and Fe, exch. K, Mg and Al+H decreased with years of cultivation; these changes were more evident in the 0-20 cm soil depth. Overall improvement of SOM and soil nutrient status was much higher in the ITAS than INAS. The levels of physical and chemical properties of soil under cacao genotypes showed a marked difference in both systems.

Trichoderma martiale sp. nov., a new endophyte from sapwood of Theobroma cacao with a potential for biological control.

PubMed

Hanada, Rogério E; de Jorge Souza, T; Pomella, Alan W V; Hebbar, K Prakash; Pereira, José O; Ismaiel, Adnan; Samuels, Gary J

2008-11-01

The new species Trichoderma martiale was isolated as an endophyte from sapwood in trunks of Theobroma cacao (cacao, Malvaceae) in Brazil. Based on sequences of translation-elongation factor 1-alpha (tef1) and RNA polymerase II subunit (rpb2) T. martiale is a close relative of, and morphologically similar to, T. viride, but differs in the production of discrete pustules on corn meal-dextrose agar (CMD) and SNA, in having a faster rate of growth, and in being a tropical endophyte. This new species was shown, in small-scale, in situ field assays, to limit black pod rot of cacao caused by Phytophthora palmivora, the cause of black pod disease.

Transposon fingerprinting using low coverage whole genome shotgun sequencing in Cacao (Theobroma cacao L.) and related species

PubMed Central

2013-01-01

Background Transposable elements (TEs) and other repetitive elements are a large and dynamically evolving part of eukaryotic genomes, especially in plants where they can account for a significant proportion of genome size. Their dynamic nature gives them the potential for use in identifying and characterizing crop germplasm. However, their repetitive nature makes them challenging to study using conventional methods of molecular biology. Next generation sequencing and new computational tools have greatly facilitated the investigation of TE variation within species and among closely related species. Results (i) We generated low-coverage Illumina whole genome shotgun sequencing reads for multiple individuals of cacao (Theobroma cacao) and related species. These reads were analysed using both an alignment/mapping approach and a de novo (graph based clustering) approach. (ii) A standard set of ultra-conserved orthologous sequences (UCOS) standardized TE data between samples and provided phylogenetic information on the relatedness of samples. (iii) The mapping approach proved highly effective within the reference species but underestimated TE abundance in interspecific comparisons relative to the de novo methods. (iv) Individual T. cacao accessions have unique patterns of TE abundance indicating that the TE composition of the genome is evolving actively within this species. (v) LTR/Gypsy elements are the most abundant, comprising c.10% of the genome. (vi) Within T. cacao the retroelement families show an order of magnitude greater sequence variability than the DNA transposon families. (vii) Theobroma grandiflorum has a similar TE composition to T. cacao, but the related genus Herrania is rather different, with LTRs making up a lower proportion of the genome, perhaps because of a massive presence (c. 20%) of distinctive low complexity satellite-like repeats in this genome. Conclusions (i) Short read alignment/mapping to reference TE contigs provides a simple and effective method of investigating intraspecific differences in TE composition. It is not appropriate for comparing repetitive elements across the species boundaries, for which de novo methods are more appropriate. (ii) Individual T. cacao accessions have unique spectra of TE composition indicating active evolution of TE abundance within this species. TE patterns could potentially be used as a “fingerprint” to identify and characterize cacao accessions. PMID:23883295

Antitumor activity against murine lymphoma L5178Y model of proteins from cacao (Theobroma cacao L.) seeds in relation with in vitro antioxidant activity.

PubMed

Preza, Ana M; Jaramillo, María E; Puebla, Ana M; Mateos, Juan C; Hernández, Rodolfo; Lugo, Eugenia

2010-10-20

Recently, proteins and peptides have become an added value to foodstuffs due to new knowledge about its structural analyses as related to antioxidant and anticancer activity. Our goal was to evaluate if protein fractions from cacao seeds show antitumor activity on lymphoma murine L5178Y model. The antioxidant activity of these fractions was also evaluated with the aim of finding a correlation with the antitumor activity. Differential extraction of proteins from unfermented and semi-fermented-dry cacao seeds was performed and characterized by SDS-PAGE and FPLC size-exclusion chromatography. Antitumor activity was evaluated against murine lymphoma L5178Y in BALB/c mice (6 × 104 cells i.p.), with a treatment oral dose of 25 mg/kg/day of each protein fraction, over a period of 15 days. Antioxidant activity was evaluated by the ABTS+ and ORAC-FL assays. Albumin, globulin and glutelin fractions from both cacao seed type were obtained by differential solubility extraction. Glutelins were the predominant fraction. In the albumin fraction, polypeptides of 42.3 and 8.5 kDa were found in native conditions, presumably in the form of two peptide chains of 21.5 kDa each one. The globulin fraction presented polypeptides of 86 and 57 kDa in unfermented cacao seed that produced the specific-cacao aroma precursors, and after fermentation the polypeptides were of 45 and 39 kDa. The glutelin fraction presented proteins >200 kDa and globulins components <100 KDa in lesser proportion. Regarding the semifermented-dry cacao seed, it was observed that the albumin fraction showed antitumoral activity, since it caused significant decreases (p < 0.05) in the ascetic fluid volume and packed cell volume, inhibiting cell growth in 59.98 ± 13.6% at 60% of the population; while the greatest antioxidant capacity due to free radical scavenging capacity was showed by the albumin and glutelin fraction in both methods assayed. This study is the first report on the biological activity of semifermented-dry cacao protein fractions with their identification, supporting the traditional use of the plant. The albumin fraction showed antitumor and free radical scavenging capacity, however both activities were not correlated. The protein fractions could be considered as source of potential antitumor peptides.

Antitumor activity against murine lymphoma L5178Y model of proteins from cacao (Theobroma cacao L.) seeds in relation with in vitro antioxidant activity

PubMed Central

2010-01-01

Background Recently, proteins and peptides have become an added value to foodstuffs due to new knowledge about its structural analyses as related to antioxidant and anticancer activity. Our goal was to evaluate if protein fractions from cacao seeds show antitumor activity on lymphoma murine L5178Y model. The antioxidant activity of these fractions was also evaluated with the aim of finding a correlation with the antitumor activity. Methods Differential extraction of proteins from unfermented and semi-fermented-dry cacao seeds was performed and characterized by SDS-PAGE and FPLC size-exclusion chromatography. Antitumor activity was evaluated against murine lymphoma L5178Y in BALB/c mice (6 × 104 cells i.p.), with a treatment oral dose of 25 mg/kg/day of each protein fraction, over a period of 15 days. Antioxidant activity was evaluated by the ABTS+ and ORAC-FL assays. Results Albumin, globulin and glutelin fractions from both cacao seed type were obtained by differential solubility extraction. Glutelins were the predominant fraction. In the albumin fraction, polypeptides of 42.3 and 8.5 kDa were found in native conditions, presumably in the form of two peptide chains of 21.5 kDa each one. The globulin fraction presented polypeptides of 86 and 57 kDa in unfermented cacao seed that produced the specific-cacao aroma precursors, and after fermentation the polypeptides were of 45 and 39 kDa. The glutelin fraction presented proteins >200 kDa and globulins components <100 KDa in lesser proportion. Regarding the semifermented-dry cacao seed, it was observed that the albumin fraction showed antitumoral activity, since it caused significant decreases (p < 0.05) in the ascetic fluid volume and packed cell volume, inhibiting cell growth in 59.98 ± 13.6% at 60% of the population; while the greatest antioxidant capacity due to free radical scavenging capacity was showed by the albumin and glutelin fraction in both methods assayed. Conclusion This study is the first report on the biological activity of semifermented-dry cacao protein fractions with their identification, supporting the traditional use of the plant. The albumin fraction showed antitumor and free radical scavenging capacity, however both activities were not correlated. The protein fractions could be considered as source of potential antitumor peptides. PMID:20961452

Cacao bean husk: an applicable bedding material in dairy free-stall barns.

PubMed

Yajima, Akira; Owada, Hisashi; Kobayashi, Suguru; Komatsu, Natsumi; Takehara, Kazuaki; Ito, Maria; Matsuda, Kazuhide; Sato, Kan; Itabashi, Hisao; Sugimura, Satoshi; Kanda, Shuhei

2017-07-01

The objectives of the study were to assess the effect of cacao bean husk as bedding material in free-stall barn on the behavior, productivity, and udder health of dairy cattle, and on the ammonia concentrations in the barn. Four different stall surfaces (no bedding, cacao bean husk, sawdust, and chopped wheat straw) were each continuously tested for a period of 1 week to determine their effects on nine lactating Holstein cows housed in the free-stall barn with rubber matting. The lying time and the milk yield were measured between d 4 and d 7. Blood samples for plasma cortisol concentration and teat swabs for bacterial counts were obtained prior to morning milking on d 7. The time-averaged gas-phase ammonia concentrations in the barn were measured between d 2 and d 7. The cows spent approximately 2 h more per day lying in the stalls when bedding was available than without bedding. The milk yield increased in the experimental periods when cows had access to bedding materials as compared to the period without bedding. The lying time was positively correlated with the milk yield. Bacterial counts on the teat ends recorded for cows housed on cacao bean husk were significantly lower than those recorded for cows housed without bedding. Ammonia concentration under cacao bean husk bedding decreased by 6%, 15%, and 21% as compared to no bedding, sawdust, and chopped wheat straw, respectively. The cortisol concentration was lowest in the period when cacao bean husk bedding was used. We observed a positive correlation between the ammonia concentrations in the barn and the plasma cortisol concentrations. Cacao bean husk is a potential alternative of conventional bedding material, such as sawdust or chopped wheat straw, with beneficial effects on udder health and ammonia concentrations in the barns.

System level analysis of cacao seed ripening reveals a sequential interplay of primary and secondary metabolism leading to polyphenol accumulation and preparation of stress resistance.

PubMed

Wang, Lei; Nägele, Thomas; Doerfler, Hannes; Fragner, Lena; Chaturvedi, Palak; Nukarinen, Ella; Bellaire, Anke; Huber, Werner; Weiszmann, Jakob; Engelmeier, Doris; Ramsak, Ziva; Gruden, Kristina; Weckwerth, Wolfram

2016-08-01

Theobroma cacao and its popular product, chocolate, are attracting attention due to potential health benefits including antioxidative effects by polyphenols, anti-depressant effects by high serotonin levels, inhibition of platelet aggregation and prevention of obesity-dependent insulin resistance. The development of cacao seeds during fruit ripening is the most crucial process for the accumulation of these compounds. In this study, we analyzed the primary and the secondary metabolome as well as the proteome during Theobroma cacao cv. Forastero seed development by applying an integrative extraction protocol. The combination of multivariate statistics and mathematical modelling revealed a complex consecutive coordination of primary and secondary metabolism and corresponding pathways. Tricarboxylic acid (TCA) cycle and aromatic amino acid metabolism dominated during the early developmental stages (stages 1 and 2; cell division and expansion phase). This was accompanied with a significant shift of proteins from phenylpropanoid metabolism to flavonoid biosynthesis. At stage 3 (reserve accumulation phase), metabolism of sucrose switched from hydrolysis into raffinose synthesis. Lipids as well as proteins involved in lipid metabolism increased whereas amino acids and N-phenylpropenoyl amino acids decreased. Purine alkaloids, polyphenols, and raffinose as well as proteins involved in abiotic and biotic stress accumulated at stage 4 (maturation phase) endowing cacao seeds the characteristic astringent taste and resistance to stress. In summary, metabolic key points of cacao seed development comprise the sequential coordination of primary metabolites, phenylpropanoid, N-phenylpropenoyl amino acid, serotonin, lipid and polyphenol metabolism thereby covering the major compound classes involved in cacao aroma and health benefits. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

Counternarcotics Assistance: U.S. Agencies Have Allotted Billions in Andean Countries, but DOD Should Improve Its Reporting of Results

DTIC Science & Technology

2012-07-01

growing areas of the world, and replaced it with licit, high-value crops such as cacao , coffee, and oil palm. The U.S. strategy seeks to replicate...producing communities transition to cultivating licit crops. Licit crops fostered by USAID programs include cacao , palm oil, and coffee. Since...include cacao , coffee, and oil palm. Counternarcotics Assistance in the Andean Region U.S. agencies use a variety of funding sources

Enhanced heavy metal immobilization in soil by grinding with addition of nanometallic Ca/CaO dispersion mixture.

PubMed

Mallampati, Srinivasa Reddy; Mitoma, Yoshiharu; Okuda, Tetsuji; Sakita, Shogo; Kakeda, Mitsunori

2012-10-01

This study investigated the use of a nanometallic Ca and CaO dispersion mixture for the immobilization of heavy metals (As, Cd, Cr and Pb) in contaminated soil. Simple grinding achieved 85-90% heavy metal immobilization, but it can be enhanced further to 98-100% by addition of a nanometallic Ca/CaO dispersion mixture produced by grinding. Observations using SEM-EDS elemental maps and semi-quantitative analysis showed that the amounts of As, Cd, Cr, and Pb measurable on the soil particle surface decrease after nanometallic Ca/CaO treatment. The leachable heavy metal concentrations were reduced after nanometallic Ca/CaO treatment to concentrations lower than the Japan soil elution standard regulatory threshold: <0.01 mg L(-1) for As, Cd, and Pb; and 0.05 mg L(-1) for Cr. Effects of soil moisture and pH on heavy metal immobilization were not strongly influenced. The most probable mechanisms for the enhancement of heavy metal immobilization capacity with nanometallic Ca/CaO treatment might be due to adsorption and entrapment of heavy metals into newly formed aggregates, thereby prompting aggregation of soil particles and enclosure/binding with Ca/CaO-associated immobile salts. Results suggest that the nanometallic Ca/CaO mixture is suitable for use in immobilization of heavy-metal-contaminated soil under normal moisture conditions. Copyright © 2012 Elsevier Ltd. All rights reserved.

Proteome analysis during pod, zygotic and somatic embryo maturation of Theobroma cacao.

PubMed

Niemenak, Nicolas; Kaiser, Edward; Maximova, Siela N; Laremore, Tatiana; Guiltinan, Mark J

2015-05-15

Two dimensional electrophoresis and nano-LC-MS were performed in order to identify alterations in protein abundance that correlate with maturation of cacao zygotic and somatic embryos. The cacao pod proteome was also characterized during development. The recently published cacao genome sequence was used to create a predicted proteolytic fragment database. Several hundred protein spots were resolved on each tissue analysis, of which 72 variable spots were subjected to MS analysis, resulting in 49 identifications. The identified proteins represent an array of functional categories, including seed storage, stress response, photosynthesis and translation factors. The seed storage protein was strongly accumulated in cacao zygotic embryos compared to their somatic counterpart. However, sucrose treatment (60 g L(-1)) allows up-regulation of storage protein in SE. A high similarity in the profiles of acidic proteins was observed in mature zygotic and somatic embryos. Differential expression in both tissues was observed in proteins having high pI. Several proteins were detected exclusively in fruit tissues, including a chitinase and a 14-3-3 protein. We also identified a novel cacao protein related to known mabinlin type sweet storage proteins. Moreover, the specific presence of thaumatin-like protein, another sweet protein, was also detected in fruit tissue. We discuss our observed correlations between protein expression profiles, developmental stage and stress responses. Copyright © 2015 Elsevier GmbH. All rights reserved.

Determination of theobromine, theophylline, and caffeine in by-products of cupuacu and cacao seeds by high-performance liquid chromatography.

PubMed

Lo Coco, F; Lanuzza, F; Micali, G; Cappellano, G

2007-01-01

Theobromine, theophylline, and caffeine are determined simultaneously by a rapid and selective reversed-phase high-performance liquid chromatography (HPLC) method with UV detection in by-products of cupuacu and cacao seeds. The determination is carried out in the raw and roasted ground cupuacu seeds and in the corresponding powders obtained after pressure treatment. The by-products of both cupuacu seeds and cacao seeds are obtained under the same technological conditions. The HPLC method uses isocratic elution with a mobile phase of methanol-water-acetic acid (80:19:1) (v/v) at a flow rate of 1 mL/min and UV absorbance detection at 275 nm. Total elution time for these analytes is less than 10 min, and the detection limit for all analytes is 0.1 mg/g. The amounts of theobromine and caffeine found in all the cupuacu samples are one or more orders of magnitude lower than those from cacao. Theophylline is found in all cacao samples except for the roasted ground paste, and it is only found in the roasted ground paste in the cupuacu samples.

Concentration of cadmium in cacao beans and its relationship with soil cadmium in southern Ecuador.

PubMed

Chavez, E; He, Z L; Stoffella, P J; Mylavarapu, R S; Li, Y C; Moyano, B; Baligar, V C

2015-11-15

Cadmium (Cd) content in cacao beans above a critical level (0.6 mg kg(-1)) has raised concerns in the consumption of cacao-based chocolate. Little is available regarding Cd concentration in soil and cacao in Ecuador. The aim of this study was to determine the status of Cd in both, soils and cacao plants, in southern Ecuador. Soil samples were collected from 19 farms at 0-5, 5-15, 15-30, and 30-50 cm depths, whereas plant samples were taken from four nearby trees. Total recoverable and extractable Cd were measured at the different soil depths. Total recoverable Cd ranged from 0.88 to 2.45 and 0.06 to 2.59, averaged 1.54 and 0.85 mg kg(-1), respectively in the surface and subsurface soils whereas the corresponding values for M3-extractable Cd were 0.08 to 1.27 and 0.02 to 0.33 with mean values of 0.40 and 0.10 mg kg(-1). Surface soil in all sampling sites had total recoverable Cd above the USEPA critical level for agricultural soils (0.43 mg kg(-1)), indicating that Cd pollution occurs. Since both total recoverable and M3-extractable Cd significantly decreased depth wise, anthropogenic activities are more likely the source of contamination. Cadmium in cacao tissues decreased in the order of beans>shell>leaves. Cadmium content in cacao beans ranged from 0.02 to 3.00, averaged 0.94 mg kg(-1), and 12 out of 19 sites had bean Cd content above the critical level. Bean Cd concentration was highly correlated with M3- or HCl-extractable Cd at both the 0-5 and 5-15 cm depths (r=0.80 and 0.82 for M3, and r=0.78 and 0.82 for HCl; P<0.01). These results indicate that accumulation of Cd in surface layers results in excessive Cd in cacao beans and M3- or HCl-extractable Cd are suitable methods for predicting available Cd in the studied soils. Copyright © 2015 Elsevier B.V. All rights reserved.

Cardiovascular Benefits of Dark Chocolate?

PubMed

Higginbotham, Erin; Taub, Pam R

2015-12-01

The use of cacao for health benefits dates back at least 3000 years. Our understanding of cacao has evolved with modern science. It is now felt based on extensive research the main health benefits of cacao stem from epicatechin, a flavanol found in cacao. The process of manufacturing dark chocolate retains epicatechin, whereas milk chocolate does not contain significant amounts of epicatechin. Thus, most of the current research studies are focused on dark chocolate. Both epidemiological and clinical studies suggest a beneficial effect of dark chocolate on blood pressure, lipids, and inflammation. Proposed mechanisms underlying these benefits include enhanced nitric oxide bioavailability and improved mitochondrial structure/function. Ultimately, further studies of this promising compound are needed to elucidate its potential for prevention and treatment of cardiovascular and metabolic diseases as well as other diseases that have underlying mechanisms of mitochondrial dysfunction and nitric oxide deficiency.

Next generation sequencing elucidates cacao badnavirus diversity and reveals the existence of more than ten viral species.

PubMed

Muller, E; Ravel, S; Agret, C; Abrokwah, F; Dzahini-Obiatey, H; Galyuon, I; Kouakou, K; Jeyaseelan, E C; Allainguillaume, J; Wetten, A

2018-01-15

Cacao swollen shoot virus is a member of the family Caulimoviridae, genus Badnavirus and is naturally transmitted to Theobroma cacao (L.) by several mealybug species. CSSV populations in West African countries are highly variable and genetically structured into several different groups based on the diversity in the first part of ORF3 which encodes the movement protein. To unravel the extent of isolate diversity and address the problems of low titer and mixed viral sequences in samples, we used Illumina MiSeq and HiSeq technology. We were able to reconstruct de novo 20 new complete genomes from cacao samples collected in the Cocoa Research Institute of Ghana (CRIG) Museum and from the field samples collected in Côte d'Ivoire or Ghana. Based on the 20% threshold of nucleotide divergence in the reverse transcriptase/ribonuclease H (RT/RNase H) region which denotes species demarcation, we conclude there exist seven new species associated with the cacao swollen shoot disease. These new species along with the three already described leads to ten, the total number of the complex of viral species associated with the disease. A sample from Sri Lanka exhibiting similar leaf symptomology to West African CSSD-affected plants was also included in the study and the corresponding sequence represents the genome of a new virus named cacao bacilliform SriLanka virus (CBSLV). Copyright © 2017 Elsevier B.V. All rights reserved.

The CACAO Method for Smoothing, Gap Filling, and Characterizing Seasonal Anomalies in Satellite Time Series

NASA Technical Reports Server (NTRS)

Verger, Aleixandre; Baret, F.; Weiss, M.; Kandasamy, S.; Vermote, E.

2013-01-01

Consistent, continuous, and long time series of global biophysical variables derived from satellite data are required for global change research. A novel climatology fitting approach called CACAO (Consistent Adjustment of the Climatology to Actual Observations) is proposed to reduce noise and fill gaps in time series by scaling and shifting the seasonal climatological patterns to the actual observations. The shift and scale CACAO parameters adjusted for each season allow quantifying shifts in the timing of seasonal phenology and inter-annual variations in magnitude as compared to the average climatology. CACAO was assessed first over simulated daily Leaf Area Index (LAI) time series with varying fractions of missing data and noise. Then, performances were analyzed over actual satellite LAI products derived from AVHRR Long-Term Data Record for the 1981-2000 period over the BELMANIP2 globally representative sample of sites. Comparison with two widely used temporal filtering methods-the asymmetric Gaussian (AG) model and the Savitzky-Golay (SG) filter as implemented in TIMESAT-revealed that CACAO achieved better performances for smoothing AVHRR time series characterized by high level of noise and frequent missing observations. The resulting smoothed time series captures well the vegetation dynamics and shows no gaps as compared to the 50-60% of still missing data after AG or SG reconstructions. Results of simulation experiments as well as confrontation with actual AVHRR time series indicate that the proposed CACAO method is more robust to noise and missing data than AG and SG methods for phenology extraction.

Application of glycerol as a foliar spray activates the defence response and enhances disease resistance of Theobroma cacao.

PubMed

Zhang, Yufan; Smith, Philip; Maximova, Siela N; Guiltinan, Mark J

2015-01-01

Previous work has implicated glycerol-3-phosphate (G3P) as a mobile inducer of systemic immunity in plants. We tested the hypothesis that the exogenous application of glycerol as a foliar spray might enhance the disease resistance of Theobroma cacao through the modulation of endogenous G3P levels. We found that exogenous application of glycerol to cacao leaves over a period of 4 days increased the endogenous level of G3P and decreased the level of oleic acid (18:1). Reactive oxygen species (ROS) were produced (a marker of defence activation) and the expression of many pathogenesis-related genes was induced. Notably, the effects of glycerol application on G3P and 18:1 fatty acid content, and gene expression levels, in cacao leaves were dosage dependent. A 100 mm glycerol spray application was sufficient to stimulate the defence response without causing any observable damage, and resulted in a significantly decreased lesion formation by the cacao pathogen Phytophthora capsici; however, a 500 mm glycerol treatment led to chlorosis and cell death. The effects of glycerol treatment on the level of 18:1 and ROS were constrained to the locally treated leaves without affecting distal tissues. The mechanism of the glycerol-mediated defence response in cacao and its potential use as part of a sustainable farming system are discussed. © 2014 BSPP AND JOHN WILEY & SONS LTD.

Mapping of a Major QTL for Ceratocystis Wilt Disease in an F1 Population of Theobroma cacao.

PubMed

Fernandes, Luciel Dos Santos; Royaert, Stefan; Corrêa, Fábio M; Mustiga, Guiliana M; Marelli, Jean-Philippe; Corrêa, Ronan X; Motamayor, Juan C

2018-01-01

Cacao is an important crop, its beans are key raw materials for the chocolate and cosmetic industries. Ceratocystis wilt of cacao (CWC) caused by Ceratocystis cacaofunesta is a lethal disease for the crop. Therefore, the selection of resistant cacao varieties is one of the viable ways to minimize losses in cacao production. In this paper, we described the identification of a major QTL associated with CWC in an F1 mapping population from a cross between a resistant, "TSH 1188," and a susceptible genotype, "CCN 51." A set of 266 trees were genotyped using 3,526 single nucleotide polymorphic markers and then multiple QTL mapping analyses were performed. Two QTLs were identified on chromosomes IV and VI. The major QTL was located at 20 cM from the top position of chromosome VI, accounting for more than 60% of the phenotypic variation. The favorable allele T1, with haplotype GTT, came from the "TSH 1188" parent. It was evident that the haplotype combination T1C2 on chromosome VI was the most significant for resistance, since 93% of resistant trees had this haplotype. The major QTL converged to a genomic region of 739.4 kb that harbored nine candidate genes, including two major classes of resistance genes, which would make them the primary candidates involved in the resistance to CWC. The haplotypes detected are now used to improve the efficiency and precision of the selection of resistant trees in cacao breeding.

Shade tree spatial structure and pod production explain frosty pod rot intensity in cacao agroforests, Costa Rica.

PubMed

Gidoin, Cynthia; Avelino, Jacques; Deheuvels, Olivier; Cilas, Christian; Bieng, Marie Ange Ngo

2014-03-01

Vegetation composition and plant spatial structure affect disease intensity through resource and microclimatic variation effects. The aim of this study was to evaluate the independent effect and relative importance of host composition and plant spatial structure variables in explaining disease intensity at the plot scale. For that purpose, frosty pod rot intensity, a disease caused by Moniliophthora roreri on cacao pods, was monitored in 36 cacao agroforests in Costa Rica in order to assess the vegetation composition and spatial structure variables conducive to the disease. Hierarchical partitioning was used to identify the most causal factors. Firstly, pod production, cacao tree density and shade tree spatial structure had significant independent effects on disease intensity. In our case study, the amount of susceptible tissue was the most relevant host composition variable for explaining disease intensity by resource dilution. Indeed, cacao tree density probably affected disease intensity more by the creation of self-shading rather than by host dilution. Lastly, only regularly distributed forest trees, and not aggregated or randomly distributed forest trees, reduced disease intensity in comparison to plots with a low forest tree density. A regular spatial structure is probably crucial to the creation of moderate and uniform shade as recommended for frosty pod rot management. As pod production is an important service expected from these agroforests, shade tree spatial structure may be a lever for integrated management of frosty pod rot in cacao agroforests.

Unique haplotypes of cacao trees as revealed by trnH-psbA chloroplast DNA

PubMed Central

Gutiérrez-López, Nidia; Ovando-Medina, Isidro; Salvador-Figueroa, Miguel; Molina-Freaner, Francisco; Avendaño-Arrazate, Carlos H.

2016-01-01

Cacao trees have been cultivated in Mesoamerica for at least 4,000 years. In this study, we analyzed sequence variation in the chloroplast DNA trnH-psbA intergenic spacer from 28 cacao trees from different farms in the Soconusco region in southern Mexico. Genetic relationships were established by two analysis approaches based on geographic origin (five populations) and genetic origin (based on a previous study). We identified six polymorphic sites, including five insertion/deletion (indels) types and one transversion. The overall nucleotide diversity was low for both approaches (geographic = 0.0032 and genetic = 0.0038). Conversely, we obtained moderate to high haplotype diversity (0.66 and 0.80) with 10 and 12 haplotypes, respectively. The common haplotype (H1) for both networks included cacao trees from all geographic locations (geographic approach) and four genetic groups (genetic approach). This common haplotype (ancient) derived a set of intermediate haplotypes and singletons interconnected by one or two mutational steps, which suggested directional selection and event purification from the expansion of narrow populations. Cacao trees from Soconusco region were grouped into one cluster without any evidence of subclustering based on AMOVA (FST = 0) and SAMOVA (FST = 0.04393) results. One population (Mazatán) showed a high haplotype frequency; thus, this population could be considered an important reservoir of genetic material. The indels located in the trnH-psbA intergenic spacer of cacao trees could be useful as markers for the development of DNA barcoding. PMID:27076998

Comparison of chocolate to cacao-free white chocolate in Parkinson's disease: a single-dose, investigator-blinded, placebo-controlled, crossover trial.

PubMed

Wolz, Martin; Schleiffer, Christine; Klingelhöfer, Lisa; Schneider, Christine; Proft, Florian; Schwanebeck, Uta; Reichmann, Heinz; Riederer, Peter; Storch, Alexander

2012-11-01

A previous questionnaire study suggests an increased chocolate consumption in Parkinson's disease (PD). The cacao ingredient contains caffeine analogues and biogenic amines, such as β-phenylethylamine, with assumed antiparkinsonian effects. We thus tested the effects of 200 g of chocolate containing 80 % of cacao on UPDRS motor score after 1 and 3 h in 26 subjects with moderate non-fluctuating PD in a mono-center, single-dose, investigator-blinded crossover study using cacao-free white chocolate as placebo comparator. At 1 h after chocolate intake, mean UPDRS motor scores were mildly decreased compared to baseline in both treatments with significant results only for dark chocolate [-1.3 (95 % CI 0.18-2.52, RMANOVA F = 4.783, p = 0.013¸ Bonferroni p = 0.021 for 1 h values)]. A 2 × 2-cross-over analysis revealed no significant differences between both treatments [-0.54 ± 0.47 (95 % CI -1.50 to 0.42), p = 0.258]. Similar results were obtained at 3 h after intake. β-phenylethylamine blood levels were unaltered. Together, chocolate did not show significant improvement over white cacao-free chocolate in PD motor function.

Effects of land-use change on community composition of tropical amphibians and reptiles in Sulawesi, Indonesia.

PubMed

Wanger, Thomas C; Iskandar, Djoko T; Motzke, Iris; Brook, Barry W; Sodhi, Navjot S; Clough, Yann; Tscharntke, Teja

2010-06-01

Little is known about the effects of anthropogenic land-use change on the amphibians and reptiles of the biodiverse tropical forests of Southeast Asia. We studied a land-use modification gradient stretching from primary forest, secondary forest, natural-shade cacao agroforest, planted-shade cacao agroforest to open areas in central Sulawesi, Indonesia. We determined species richness, abundance, turnover, and community composition in all habitat types and related these to environmental correlates, such as canopy heterogeneity and thickness of leaf litter. Amphibian species richness decreased systematically along the land-use modification gradient, but reptile richness and abundance peaked in natural-shade cacao agroforests. Species richness and abundance patterns across the disturbance gradient were best explained by canopy cover and leaf-litter thickness in amphibians and by canopy heterogeneity and cover in reptiles. Amphibians were more severely affected by forest disturbance in Sulawesi than reptiles. Heterogeneous canopy cover and thick leaf litter should be maintained in cacao plantations to facilitate the conservation value for both groups. For long-term and sustainable use of plantations, pruned shade trees should be permanently kept to allow rejuvenation of cacao and, thus, to prevent repeated forest encroachment.

Cacao bean husk: an applicable bedding material in dairy free-stall barns

PubMed Central

Yajima, Akira; Owada, Hisashi; Kobayashi, Suguru; Komatsu, Natsumi; Takehara, Kazuaki; Ito, Maria; Matsuda, Kazuhide; Sato, Kan; Itabashi, Hisao; Sugimura, Satoshi; Kanda, Shuhei

2017-01-01

Objective The objectives of the study were to assess the effect of cacao bean husk as bedding material in free-stall barn on the behavior, productivity, and udder health of dairy cattle, and on the ammonia concentrations in the barn. Methods Four different stall surfaces (no bedding, cacao bean husk, sawdust, and chopped wheat straw) were each continuously tested for a period of 1 week to determine their effects on nine lactating Holstein cows housed in the free-stall barn with rubber matting. The lying time and the milk yield were measured between d 4 and d 7. Blood samples for plasma cortisol concentration and teat swabs for bacterial counts were obtained prior to morning milking on d 7. The time-averaged gas-phase ammonia concentrations in the barn were measured between d 2 and d 7. Results The cows spent approximately 2 h more per day lying in the stalls when bedding was available than without bedding. The milk yield increased in the experimental periods when cows had access to bedding materials as compared to the period without bedding. The lying time was positively correlated with the milk yield. Bacterial counts on the teat ends recorded for cows housed on cacao bean husk were significantly lower than those recorded for cows housed without bedding. Ammonia concentration under cacao bean husk bedding decreased by 6%, 15%, and 21% as compared to no bedding, sawdust, and chopped wheat straw, respectively. The cortisol concentration was lowest in the period when cacao bean husk bedding was used. We observed a positive correlation between the ammonia concentrations in the barn and the plasma cortisol concentrations. Conclusion Cacao bean husk is a potential alternative of conventional bedding material, such as sawdust or chopped wheat straw, with beneficial effects on udder health and ammonia concentrations in the barns. PMID:28002931

The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca(2+) and Mg(2+) dependent-DNase activity and antifungal action on Moniliophthora perniciosa.

PubMed

Pereira Menezes, Sara; de Andrade Silva, Edson Mario; Matos Lima, Eline; Oliveira de Sousa, Aurizângela; Silva Andrade, Bruno; Santos Lima Lemos, Livia; Peres Gramacho, Karina; da Silva Gesteira, Abelmon; Pirovani, Carlos Priminho; Micheli, Fabienne

2014-06-11

The production and accumulation of pathogenesis-related proteins (PR proteins) in plants in response to biotic or abiotic stresses is well known and is considered as a crucial mechanism for plant defense. A pathogenesis-related protein 4 cDNA was identified from a cacao-Moniliophthora perniciosa interaction cDNA library and named TcPR-4b. TcPR-4b presents a Barwin domain with six conserved cysteine residues, but lacks the chitin-binding site. Molecular modeling of TcPR-4b confirmed the importance of the cysteine residues to maintain the protein structure, and of several conserved amino acids for the catalytic activity. In the cacao genome, TcPR-4b belonged to a small multigene family organized mainly on chromosome 5. TcPR-4b RT-qPCR analysis in resistant and susceptible cacao plants infected by M. perniciosa showed an increase of expression at 48 hours after infection (hai) in both cacao genotypes. After the initial stage (24-72 hai), the TcPR-4b expression was observed at all times in the resistant genotypes, while in the susceptible one the expression was concentrated at the final stages of infection (45-90 days after infection). The recombinant TcPR-4b protein showed RNase, and bivalent ions dependent-DNase activity, but no chitinase activity. Moreover, TcPR-4b presented antifungal action against M. perniciosa, and the reduction of M. perniciosa survival was related to ROS production in fungal hyphae. To our knowledge, this is the first report of a PR-4 showing simultaneously RNase, DNase and antifungal properties, but no chitinase activity. Moreover, we showed that the antifungal activity of TcPR-4b is directly related to RNase function. In cacao, TcPR-4b nuclease activities may be related to the establishment and maintenance of resistance, and to the PCD mechanism, in resistant and susceptible cacao genotypes, respectively.

Entomofauna Associated with Agroforestry Systems of Timber Species and Cacao in the Southern Region of the Maracaibo Lake Basin (Mérida, Venezuela).

PubMed

Mazón, Marina; Sánchez-Angarita, Daniel; Díaz, Francisco A; Gutiérrez, Néstor; Jaimez, Ramón

2018-04-20

Agroforestry systems are environment-friendly production systems which help to preserve biodiversity while providing people with a way of earning a living. Cacao is a historically important crop in Venezuela that traditionally has been produced in agroforestry systems. However, few studies have evaluated how different trees used in those systems affect the dynamics and abundance of insects. The present study evaluated the entomofauna assemblages associated with different combinations of four timber-yielding trees and four Criollo cacao cultivars established in a lowland tropical ecosystem in Venezuela. A randomized block design with two replicates was used, each block having 16 plots which included all 16 possible combinations of four native timber trees ( Cordia thaisiana , Cedrela odorata , Swietenia macrophylla , and Tabebuia rosea ) and four Criollo cacao cultivars (Porcelana, Guasare, Lobatera and Criollo Merideño). Insects were collected with yellow pan traps and sorted to order. Coleoptera and parasitoid Hymenoptera were determined to the family level. In total, 49,538 individuals of seven orders were collected, with Hymenoptera, Diptera, and Hemiptera being the most abundant, although only Lepidoptera and Coleoptera abundances were significantly influenced by the timber tree species. Twenty-three families of parasitoid Hymenoptera and 26 of Coleoptera were found. Significant differences in insects’ assemblages were found both in parasitoid Hymenoptera and Coleoptera families associated to every shade tree, with the families Eulophidae and Lycidae being indicators for Cordia , and Chalcididae for Swietenia . The entomofauna relationship with the cacao cultivar was barely significant, although Scydmaenidae and Scarabaeidae were indicators for Lobatera and Merideño, respectively. No significant effects were found for interaction with cacao cultivars and native trees. We concluded that the particular insect assemblages found in Cedrela odorata and Cordia thaisiana , together with their high growing rates, make these two species an optimal choice for cacao agroforestry systems.

Proanthocyanidin synthesis in Theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase

PubMed Central

2013-01-01

Background The proanthocyanidins (PAs), a subgroup of flavonoids, accumulate to levels of approximately 10% total dry weight of cacao seeds. PAs have been associated with human health benefits and also play important roles in pest and disease defense throughout the plant. Results To dissect the genetic basis of PA biosynthetic pathway in cacao (Theobroma cacao), we have isolated three genes encoding key PA synthesis enzymes, anthocyanidin synthase (ANS), anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR). We measured the expression levels of TcANR, TcANS and TcLAR and PA content in cacao leaves, flowers, pod exocarp and seeds. In all tissues examined, all three genes were abundantly expressed and well correlated with PA accumulation levels, suggesting their active roles in PA synthesis. Overexpression of TcANR in an Arabidopsis ban mutant complemented the PA deficient phenotype in seeds and resulted in reduced anthocyanidin levels in hypocotyls. Overexpression of TcANS in tobacco resulted in increased content of both anthocyanidins and PAs in flower petals. Overexpression of TcANS in an Arabidopsis ldox mutant complemented its PA deficient phenotype in seeds. Recombinant TcLAR protein converted leucoanthocyanidin to catechin in vitro. Transgenic tobacco overexpressing TcLAR had decreased amounts of anthocyanidins and increased PAs. Overexpressing TcLAR in Arabidopsis ldox mutant also resulted in elevated synthesis of not only catechin but also epicatechin. Conclusion Our results confirm the in vivo function of cacao ANS and ANR predicted based on sequence homology to previously characterized enzymes from other species. In addition, our results provide a clear functional analysis of a LAR gene in vivo. PMID:24308601

Association mapping of seed and disease resistance traits in Theobroma cacao L.

PubMed

Motilal, Lambert A; Zhang, Dapeng; Mischke, Sue; Meinhardt, Lyndel W; Boccara, Michel; Fouet, Olivier; Lanaud, Claire; Umaharan, Pathmanathan

2016-12-01

Microsatellite and single nucleotide polymorphism markers that could be used in marker assisted breeding of cacao were identified for number of filled seeds, black pod resistance and witches' broom disease resistance. An association mapping approach was employed to identify markers for seed number and resistance to black pod and witches' broom disease (WBD) in cacao (Theobroma cacao L.). Ninety-five microsatellites (SSRs) and 775 single nucleotide polymorphisms (SNPs) were assessed on 483 unique trees in the International Cocoa Genebank Trinidad (ICGT). Linkage disequilibrium (LD) and association mapping studies were conducted to identify markers to tag the phenotypic traits. Decay of LD occurred over an average 9.3 cM for chromosomes 1-9 and 2.5 cM for chromosome 10. Marker/trait associations were generally identified based on general linear models (GLMs) that incorporated principal components from molecular information on relatedness factor. Seven markers (mTcCIR 8, 66, 126, 212; TcSNP368, 697, 1370) on chromosomes 1 and 9 were identified for number of filled seeds (NSEED). A single marker was found for black pod resistance (mTcCIR280) on chromosome 3, whereas six markers on chromosomes 4, 5, 6, 8, and 10 were detected for WBD (mTcCIR91, 183; TcSNP375, 720, 1230 and 1374). It is expected that this association mapping study in cacao would contribute to the knowledge of the genetic determinism of cocoa traits and that the markers identified herein would prove useful in marker assisted breeding of cacao.

Proanthocyanidin synthesis in Theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase.

PubMed

Liu, Yi; Shi, Zi; Maximova, Siela; Payne, Mark J; Guiltinan, Mark J

2013-12-05

The proanthocyanidins (PAs), a subgroup of flavonoids, accumulate to levels of approximately 10% total dry weight of cacao seeds. PAs have been associated with human health benefits and also play important roles in pest and disease defense throughout the plant. To dissect the genetic basis of PA biosynthetic pathway in cacao (Theobroma cacao), we have isolated three genes encoding key PA synthesis enzymes, anthocyanidin synthase (ANS), anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR). We measured the expression levels of TcANR, TcANS and TcLAR and PA content in cacao leaves, flowers, pod exocarp and seeds. In all tissues examined, all three genes were abundantly expressed and well correlated with PA accumulation levels, suggesting their active roles in PA synthesis. Overexpression of TcANR in an Arabidopsis ban mutant complemented the PA deficient phenotype in seeds and resulted in reduced anthocyanidin levels in hypocotyls. Overexpression of TcANS in tobacco resulted in increased content of both anthocyanidins and PAs in flower petals. Overexpression of TcANS in an Arabidopsis ldox mutant complemented its PA deficient phenotype in seeds. Recombinant TcLAR protein converted leucoanthocyanidin to catechin in vitro. Transgenic tobacco overexpressing TcLAR had decreased amounts of anthocyanidins and increased PAs. Overexpressing TcLAR in Arabidopsis ldox mutant also resulted in elevated synthesis of not only catechin but also epicatechin. Our results confirm the in vivo function of cacao ANS and ANR predicted based on sequence homology to previously characterized enzymes from other species. In addition, our results provide a clear functional analysis of a LAR gene in vivo.

Two Theobroma cacao genotypes with contrasting pathogen tolerance show aberrant transcriptional and ROS responses after salicylic acid treatment.

PubMed

Fister, Andrew S; O'Neil, Shawn T; Shi, Zi; Zhang, Yufan; Tyler, Brett M; Guiltinan, Mark J; Maximova, Siela N

2015-10-01

Understanding the genetic basis of pathogen susceptibility in various crop plants is crucial to increasing the stability of food, feed, and fuel production. Varietal differences in defence responses provide insights into the mechanisms of resistance and are a key resource for plant breeders. To explore the role of salicylic acid in the regulation of defence in cacao, we demonstrated that SA treatment decreased susceptibility to a pod rot pathogen, Phytophthora tropicalis in two genotypes, Scavina 6 and Imperial College Selection 1, which differ in their resistance to several agriculturally important pathogens. Transient overexpression of TcNPR1, a major transcriptional regulator of the SA-dependent plant immune system, also increased pathogen tolerance in cacao leaves. To explore further the genetic basis of resistance in cacao, we used microarrays to measure gene expression profiles after salicylic acid (SA) treatment in these two cacao genotypes. The two genotypes displayed distinct transcriptional responses to SA. Unexpectedly, the expression profile of the susceptible genotype ICS1 included a larger number of pathogenesis-related genes that were induced by SA at 24h after treatment, whereas genes encoding many chloroplast and mitochondrial proteins implicated in reactive oxygen species production were up-regulated in the resistant genotype, Sca6. Sca6 accumulated significantly more superoxide at 24h after treatment of leaves with SA. These experiments revealed critical insights regarding the molecular differences between cacao varieties, which will allow a better understanding of defence mechanisms to help guide breeding programmes. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Two Theobroma cacao genotypes with contrasting pathogen tolerance show aberrant transcriptional and ROS responses after salicylic acid treatment

PubMed Central

Fister, Andrew S.; O’Neil, Shawn T.; Shi, Zi; Zhang, Yufan; Tyler, Brett M.; Guiltinan, Mark J.; Maximova, Siela N.

2015-01-01

Understanding the genetic basis of pathogen susceptibility in various crop plants is crucial to increasing the stability of food, feed, and fuel production. Varietal differences in defence responses provide insights into the mechanisms of resistance and are a key resource for plant breeders. To explore the role of salicylic acid in the regulation of defence in cacao, we demonstrated that SA treatment decreased susceptibility to a pod rot pathogen, Phytophthora tropicalis in two genotypes, Scavina 6 and Imperial College Selection 1, which differ in their resistance to several agriculturally important pathogens. Transient overexpression of TcNPR1, a major transcriptional regulator of the SA-dependent plant immune system, also increased pathogen tolerance in cacao leaves. To explore further the genetic basis of resistance in cacao, we used microarrays to measure gene expression profiles after salicylic acid (SA) treatment in these two cacao genotypes. The two genotypes displayed distinct transcriptional responses to SA. Unexpectedly, the expression profile of the susceptible genotype ICS1 included a larger number of pathogenesis-related genes that were induced by SA at 24h after treatment, whereas genes encoding many chloroplast and mitochondrial proteins implicated in reactive oxygen species production were up-regulated in the resistant genotype, Sca6. Sca6 accumulated significantly more superoxide at 24h after treatment of leaves with SA. These experiments revealed critical insights regarding the molecular differences between cacao varieties, which will allow a better understanding of defence mechanisms to help guide breeding programmes. PMID:26163705

Mapping of a Major QTL for Ceratocystis Wilt Disease in an F1 Population of Theobroma cacao

PubMed Central

Fernandes, Luciel dos Santos; Royaert, Stefan; Corrêa, Fábio M.; Mustiga, Guiliana M.; Marelli, Jean-Philippe; Corrêa, Ronan X.; Motamayor, Juan C.

2018-01-01

Cacao is an important crop, its beans are key raw materials for the chocolate and cosmetic industries. Ceratocystis wilt of cacao (CWC) caused by Ceratocystis cacaofunesta is a lethal disease for the crop. Therefore, the selection of resistant cacao varieties is one of the viable ways to minimize losses in cacao production. In this paper, we described the identification of a major QTL associated with CWC in an F1 mapping population from a cross between a resistant, “TSH 1188,” and a susceptible genotype, “CCN 51.” A set of 266 trees were genotyped using 3,526 single nucleotide polymorphic markers and then multiple QTL mapping analyses were performed. Two QTLs were identified on chromosomes IV and VI. The major QTL was located at 20 cM from the top position of chromosome VI, accounting for more than 60% of the phenotypic variation. The favorable allele T1, with haplotype GTT, came from the “TSH 1188” parent. It was evident that the haplotype combination T1C2 on chromosome VI was the most significant for resistance, since 93% of resistant trees had this haplotype. The major QTL converged to a genomic region of 739.4 kb that harbored nine candidate genes, including two major classes of resistance genes, which would make them the primary candidates involved in the resistance to CWC. The haplotypes detected are now used to improve the efficiency and precision of the selection of resistant trees in cacao breeding. PMID:29491879

Dinitrogen fixation by legume shade trees and direct transfer of fixed N to associated cacao in a tropical agroforestry system.

PubMed

Nygren, Pekka; Leblanc, Humberto A

2015-02-01

Natural abundance of (15)N (δ (15)N) was determined in bulk soil, rhizospheric soil and vegetation in an organically managed cacao (Theobroma cacao L.) plantation with Inga edulis Mart. legume trees (inga) as the principal shade for studying the nitrogen (N) cycle in the system. Cacao without contact with legumes in an adjacent plantation was used as the reference for N2 fixation and direct N transfer calculations. Bulk and rhizospheric soils contained 72 and 20%, respectively, of whole- system N. No vegetation effect on δ (15)N in rhizospheric soil was detected, probably due to the high native soil N pool. Fine roots of the cacaos associated with inga contained ∼35% of N fixed from the atmosphere (Nf) out of the total N. Leaves of all species had significantly higher δ (15)N than fine roots. Twenty percent of system Nf was found in cacao suggesting direct N transfer from inga via a common mycelial network of mycorrhizal fungi or recycling of N-rich root exudates of inga. Inga had accumulated 98 kg [Nf] ha(-1) during the 14-year history of the plantation. The conservative estimate of current N2 fixation rate was 41 kg [Nf] ha(-1) year(-1) based on inga biomass only and 50 kg [Nf] ha(-1) year(-1) based on inga and associated trees. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Phytophthora megakarya and P. palmivora, Causal Agents of Black Pod Rot, Induce Similar Plant Defense Responses Late during Infection of Susceptible Cacao Pods

PubMed Central

Ali, Shahin S.; Shao, Jonathan; Lary, David J.; Strem, Mary D.; Meinhardt, Lyndel W.; Bailey, Bryan A.

2017-01-01

Phytophthora megakarya (Pmeg) and Phytophthora palmivora (Ppal) cause black pod rot of Theobroma cacao L. (cacao). Of these two clade 4 species, Pmeg is more virulent and is displacing Ppal in many cacao production areas in Africa. Symptoms and species specific sporangia production were compared when the two species were co-inoculated onto pod pieces in staggered 24 h time intervals. Pmeg sporangia were predominantly recovered from pod pieces with unwounded surfaces even when inoculated 24 h after Ppal. On wounded surfaces, sporangia of Ppal were predominantly recovered if the two species were simultaneously applied or Ppal was applied first but not if Pmeg was applied first. Pmeg demonstrated an advantage over Ppal when infecting un-wounded surfaces while Ppal had the advantage when infecting wounded surfaces. RNA-Seq was carried out on RNA isolated from control and Pmeg and Ppal infected pod pieces 3 days post inoculation to assess their abilities to alter/suppress cacao defense. Expression of 4,482 and 5,264 cacao genes was altered after Pmeg and Ppal infection, respectively, with most genes responding to both species. Neural network self-organizing map analyses separated the cacao RNA-Seq gene expression profiles into 24 classes, 6 of which were largely induced in response to infection. Using KEGG analysis, subsets of genes composing interrelated pathways leading to phenylpropanoid biosynthesis, ethylene and jasmonic acid biosynthesis and action, plant defense signal transduction, and endocytosis showed induction in response to infection. A large subset of genes encoding putative Pr-proteins also showed differential expression in response to infection. A subset of 36 cacao genes was used to validate the RNA-Seq expression data and compare infection induced gene expression patterns in leaves and wounded and unwounded pod husks. Expression patterns between RNA-Seq and RT-qPCR were generally reproducible. The level and timing of altered gene expression was influenced by the tissues studied and by wounding. Although, in these susceptible interactions gene expression patterns were similar, some genes did show differential expression in a Phytophthora species dependent manner. The biggest difference was the more intense changes in expression in Ppal inoculated wounded pod pieces further demonstrating its rapid progression when penetrating through wounds. PMID:28261234

Automatic start control for a three-phase electric motor using infrared sensors

NASA Astrophysics Data System (ADS)

Echenique Lima, Mario; Ramírez Arenas, Francisco; Rodríguez Pedroza, Griselda

2006-02-01

We introduce equipment for the automatic activation of a three-phase electric motor (1Hp, 3A, 240V AC) using 2 infrared sensors monitored by a Microchip microcontroller PIC16F62x@4Mhz for the control of a filling system. This project was carried out to Fabrica de Chocolates y Dulces Costanzo, where the automatization of cacao grain supply was required for a machine in charge of cleaning the cacao from its rind. This process demanded the monitoring of the filling level to avoid the spill of toasted cacao.

Cacao usage by the earliest Maya civilization.

PubMed

Hurst, W Jeffrey; Tarka, Stanley M; Powis, Terry G; Valdez, Fred; Hester, Thomas R

2002-07-18

The Maya archaeological site at Colha in northern Belize, Central America, has yielded several spouted ceramic vessels that contain residues from the preparation of food and beverages. Here we analyse dry residue samples by using high-performance liquid chromatography coupled to atmospheric-pressure chemical-ionization mass spectrometry, and show that chocolate (Theobroma cacao) was consumed by the Preclassic Maya as early as 600 bc, pushing back the earliest chemical evidence of cacao use by some 1,000 years. Our application of this new and highly sensitive analytical technique could be extended to the identification of other ancient foods and beverages.

Cacao Intensification in Sulawesi: A Green Prosperity Model Project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moriarty, K.; Elchinger, M.; Hill, G.

2014-09-01

NREL conducted eight model projects for Millennium Challenge Corporation's (MCC) Compact with Indonesia. Green Prosperity, the largest project of the Compact, seeks to address critical constraints to economic growth while supporting the Government of Indonesia's commitment to a more sustainable, less carbon-intensive future. This study evaluates techniques to improve cacao farming in Sulawesi Indonesia with an emphasis on Farmer Field Schools and Cocoa Development Centers to educate farmers and for train the trainer programs. The study estimates the economic viability of cacao farming if smallholder implement techniques to increase yield as well as social and environmental impacts of the project.

Studies on the stability of acrylamide in food during storage.

PubMed

Hoenicke, Katrin; Gatermann, Robert

2005-01-01

Acrylamide levels in a variety of food samples were analyzed before and after 3 months of storage at 10 degrees-12 degrees C. The analysis was performed by liquid chromatography tandem mass spectrometry (LC/MS/MS) using deuterium-labeled acrylamide as internal standard. Acrylamide was stable in most matrixes (cookies, cornflakes, crispbread, raw sugar, potato crisps, peanuts) over time. However, slight decreases were determined for dietary biscuits (83-89%) and for licorice confection (82%). For coffee and cacao powder, a significant decrease occurred during storage for 3 or 6 months, respectively. Acrylamide concentrations dropped from 305 to 210 microg/kg in coffee and from 265 to 180 microg/kg in cacao powder. On the contrary, acrylamide remained stable in soluble coffee as well as in coffee substitutes. Reactions of acrylamide with SH group-containing substances were assumed as the cause for acrylamide degradation in coffee and cacao. Spiking experiments with acrylamide revealed that acrylamide concentrations remained stable in baby food, cola, and beer; however, recovery levels dropped in milk powder (71%), sulfurized apricot (53%), and cacao powder (17%). These observations suggest that variations in the acrylamide content of food, especially in coffee and cacao, can vary depending on the storage time because special food constituents and/or reaction products can affect the levels.

Comparative proteomic analysis of early somatic and zygotic embryogenesis in Theobroma cacao L.

PubMed

Noah, Alexandre Mboene; Niemenak, Nicolas; Sunderhaus, Stephanie; Haase, Christin; Omokolo, Denis Ndoumou; Winkelmann, Traud; Braun, Hans-Peter

2013-01-14

Somatic embryogenesis can efficiently foster the propagation of Theobroma cacao, but the poor quality of resulted plantlet hinders the use of this technique in the commercial scale. The current study has been initiated to systematically compare the physiological mechanisms underlying somatic and zygotic embryogenesis in T. cacao on the proteome level. About 1000 protein spots per fraction could be separated by two-dimensional isoelectric focusing/SDS PAGE. More than 50 of the protein spots clearly differed in abundance between zygotic and somatic embryos: 33 proteins spots were at least 3-fold higher in abundance in zygotic embryos and 20 in somatic embryos. Analyses of these protein spots differing in volume by mass spectrometry resulted in the identification of 68 distinct proteins. Many of the identified proteins are involved in genetic information processing (21 proteins), carbohydrate metabolism (11 proteins) and stress response (7 proteins). Somatic embryos especially displayed many stress related proteins, few enzymes involved in storage compound synthesis and an exceptional high abundance of endopeptidase inhibitors. Phosphoenolpyruvate carboxylase, which was accumulated more than 3-fold higher in zygotic embryos, represents a prominent enzyme in the storage compound metabolism in cacao seeds. Implications on the improvement of somatic embryogenesis in cacao are discussed. Copyright © 2012 Elsevier B.V. All rights reserved.

[Transmission of cutaneous leishmaniasis associated with cacao (Theobroma cacao) plantations in Tabasco].

PubMed

Carrada Figueroa, Georgina Del Carmen; Leal Ascencio, Víctor Javier; Jiménez Sastré, Alejandro; López Álvarez, Jorge

2014-01-01

Tabasco is the Mexican state that reported the highest number (37.4%) of patients with leishmaniasis during 1990-2011. Close to 90% of these patients lived in Chontalpa, where the municipality of Cunduacán accounted for the majority of the cases. One of the characteristics of this region is that houses are located within cacao plantations. To determine if cacao plantations are a risk factor for leishmaniasis transmission in locations of Cunduacán, Tabasco. We performed an analytical and retrospective study of 115 locations in Cunduacán, analyzing the number of localities with or without patients with leishmaniasis registered between 2000-2011 and, additionally, if they had cacao plantations, using a map where different crops were georeferenced. We measured the magnitude of the association (odds ratio, 95% CI). During the period 2000-2011, cases of leishmaniasis were reported in 77 (67.0%) Cunduacán locations, of these, 55 (71.4%) had cocoa plantations, five (6.5%) of banana, five (6.5%) of cane, and 12 (15.6%) had no crops georeferenced. We found that cocoa crops are a risk factor for the transmission of leishmaniasis (OR: 3.438; 95% CI: 1,526-7,742). The probability of transmission of leishmaniasis in areas with cocoa crops is greater than in communities without this crop.

Anti-inflammatory activity of Theobroma cacao L. stem bark ethanol extract and its fractions in experimental models.

PubMed

Oyeleke, Sabitiu A; Ajayi, Abayomi M; Umukoro, Solomon; Aderibigbe, A O; Ademowo, Olusegun George

2018-08-10

The stem bark of Theobroma cacao L. have been used for the treatment of inflammation, toothache, measles and malaria in ethnomedicine. However, the anti-inflammatory activity of Theobroma cacao stem bark has not been fully elucidated. The anti-inflammatory activity of Theobroma cacao stem bark ethanol extract and its fractions was investigated in this study. The anti-inflammatory effect of ethanol extract of Theobroma cacao stem bark (EETc) and its dichloromethane (DCMF), ethylacetate (EAF) and aqueous (AQF) fractions was investigated in erythrocytes membrane stabilizing assay and carrageenan-induced paw oedema. The anti-inflammatory activity of the EAF and EETc was investigated in carrageenan induced-granuloma air pouch models. The extract and fractions showed significant membrane stabilizing action on rat erythrocytes cell membrane. The oral administration of DCMF, EAF and AQF (250 mg/kg) significantly inhibited paw oedema induced by carrageenan (41.3%, 55.0% and 45.0%, respectively) compared to control group. The EAF (62.5, 125 and 250 mg/kg) and EETc (250 mg/kg) significantly inhibited exudates formation in carrageenan air pouch by (63.8, 71.5, 74.5, 64.3%) at 24 h and by (69.4%, 75.7%, 77.1% and 68.4%) at 72 h respectively. The EETc and EAF significantly reduced neutrophil counts, protein, nitrite, Tumor necrosis factor (TNF-α) and malondialdehyde (MDA) but increased reduced glutathione (GSH) levels compared to control in pouch exudates. The HPLC fingerprint of EAF revealed presence of caffeic acid, rutin, ferulic acid and morin. Ethanol extract of Theobroma cacao and its ethylacetate fraction demonstrated anti-inflammatory activity partly by reducing neutrophil migration and inflammatory mediator production. Copyright © 2018 Elsevier B.V. All rights reserved.

The Theobroma cacao B3 domain transcription factor TcLEC2 plays a duel role in control of embryo development and maturation.

PubMed

Zhang, Yufan; Clemens, Adam; Maximova, Siela N; Guiltinan, Mark J

2014-04-24

The Arabidopsis thaliana LEC2 gene encodes a B3 domain transcription factor, which plays critical roles during both zygotic and somatic embryogenesis. LEC2 exerts significant impacts on determining embryogenic potential and various metabolic processes through a complicated genetic regulatory network. An ortholog of the Arabidopsis Leafy Cotyledon 2 gene (AtLEC2) was characterized in Theobroma cacao (TcLEC2). TcLEC2 encodes a B3 domain transcription factor preferentially expressed during early and late zygotic embryo development. The expression of TcLEC2 was higher in dedifferentiated cells competent for somatic embryogenesis (embryogenic calli), compared to non-embryogenic calli. Transient overexpression of TcLEC2 in immature zygotic embryos resulted in changes in gene expression profiles and fatty acid composition. Ectopic expression of TcLEC2 in cacao leaves changed the expression levels of several seed related genes. The overexpression of TcLEC2 in cacao explants greatly increased the frequency of regeneration of stably transformed somatic embryos. TcLEC2 overexpressing cotyledon explants exhibited a very high level of embryogenic competency and when cultured on hormone free medium, exhibited an iterative embryogenic chain-reaction. Our study revealed essential roles of TcLEC2 during both zygotic and somatic embryo development. Collectively, our evidence supports the conclusion that TcLEC2 is a functional ortholog of AtLEC2 and that it is involved in similar genetic regulatory networks during cacao somatic embryogenesis. To our knowledge, this is the first detailed report of the functional analysis of a LEC2 ortholog in a species other then Arabidopsis. TcLEC2 could potentially be used as a biomarker for the improvement of the SE process and screen for elite varieties in cacao germplasm.

Molecular and biochemical characterisation of two aspartic proteinases TcAP1 and TcAP2 from Theobroma cacao seeds.

PubMed

Laloi, Maryse; McCarthy, James; Morandi, Olivia; Gysler, Christof; Bucheli, Peter

2002-09-01

Aspartic proteinase (EC 3.4.23) activity plays a pivotal role in the degradation of Theobroma cacao L. seed proteins during the fermentation step of cacao bean processing. Therefore, this enzyme is believed to be critical for the formation of the peptide and amino acid cocoa flavor precursors that occurs during fermentation. Using cDNA cloning and northern blot analysis, we show here that there are at least two distinct aspartic proteinase genes ( TcAP1 and TcAP2) expressed during cacao seed development. Both genes are expressed early during seed development and their mRNA levels decrease towards the end of seed maturation. TcAP2 is expressed at a much higher level than TcAP1, although the expression of TcAP1 increases slightly during germination. The proteins encoded by TcAP1 and TcAP2 are relatively different from each other (73% identity). This, and the fact that the two corresponding genes have different expression patterns, suggests that the TcAP1 and TcAP2 proteins may have different functions in the maturing seeds and during germination. Because the TcAP2 gene is expressed at a much higher level during seed development than TcAP1, it is likely that the TcAP2 protein is primarily responsible for the majority of the industrially important protein hydrolysis that occurs during cacao bean fermentation. Finally, TcAP2 has been functionally expressed in the yeast Yarrowia lipolytica. The secreted recombinant protein is able to hydrolyse bovine haemoglobin at acidic pH and is sensitive to pepstatin A, confirming that TcAP2 encodes an aspartic proteinase, and strongly suggests that this gene encodes the well-characterized aspartic proteinase of mature cacao seeds.

Breeding for disease resistance in cacao

USDA-ARS?s Scientific Manuscript database

Cacao production must increase in order to meet the projected rise in the demand for chocolate. Approximately one-third of global production is lost annually to diseases and insects. Four diseases account for the greatest losses worldwide: black pod, caused by four Phytophthora spp; witches’ broom...

Cacao diseases: A history of old enemies and new encounters

USDA-ARS?s Scientific Manuscript database

This book reviews the current knowledge of cacao pathogens and their management methods. Topics discussed include the history, biology, and genetic diversity of Moniliophthora (causing witches’ broom and frosty pod rot) and Phytophthora species (causing black pod rot) that cause diseases resulting i...

Resistance gene homologues in Theobroma cacao as useful genetic markers.

PubMed

Kuhn, D N; Heath, M; Wisser, R J; Meerow, A; Brown, J S; Lopes, U; Schnell, R J

2003-07-01

Resistance gene homologue (RGH) sequences have been developed into useful genetic markers for marker-assisted selection (MAS) of disease resistant Theobroma cacao. A plasmid library of amplified fragments was created from seven different cultivars of cacao. Over 600 cloned recombinant amplicons were evaluated. From these, 74 unique RGHs were identified that could be placed into 11 categories based on sequence analysis. Primers specific to each category were designed. The primers specific for a single RGH category amplified fragments of equal length from the seven different cultivars used to create the library. However, these fragments exhibited single-strand conformational polymorphism (SSCP), which allowed us to map six of the RGH categories in an F(2) population of T. cacao. RGHs 1, 4 and 5 were in the same linkage group, with RGH 4 and 5 separated by less than 4 cM. As SSCP can be efficiently performed on our automated sequencer, we have developed a convenient and rapid high throughput assay for RGH alleles.

The causal agents of witches' broom and frosty pod rot of cacao (chocolate, Theobroma cacao) form a new lineage of Marasmiaceae.

PubMed

Aime, M C; Phillips-Mora, W

2005-01-01

The two most devastating diseases of cacao (Theobroma cacao)--the source of chocolate--in tropical America are caused by the fungi Crinipellis perniciosa (witches' broom disease) and Moniliophthora roreri (frosty pod rot or moniliasis disease). Despite the agricultural, socio-economic and environmental impact of these fungi, most aspects of their life cycles are unknown, and the phylogenetic relationships of M. roreri have yet to be conclusively established. In this paper, extensive phylogenetic analyses of five nuclear gene regions (28S rDNA, 18S rDNA, ITS, RPB1, and EF1-alpha) confirm that C. perniciosa and M. roreri are sister taxa that belong in the Marasmiaceae (euagarics). Furthermore, these taxa form part of a separate and distinct lineage within the family. This lineage includes the biotrophic fungi Moniliophthora perniciosa comb. nov. and M. roreri, as well as one undescribed endophytic species. The sister genera to Moniliophthora are Marasmius, Crinipellis and Chaetocalathus, which consist mainly of saprotrophic litter fungi.

Development of a marker assisted selection program for cacao.

PubMed

Schnell, R J; Kuhn, D N; Brown, J S; Olano, C T; Phillips-Mora, W; Amores, F M; Motamayor, J C

2007-12-01

ABSTRACT Production of cacao in tropical America has been severely affected by fungal pathogens causing diseases known as witches' broom (WB, caused by Moniliophthora perniciosa), frosty pod (FP, caused by M. roreri) and black pod (BP, caused by Phytophthora spp.). BP is pan-tropical and causes losses in all producing areas. WB is found in South America and parts of the Caribbean, while FP is found in Central America and parts of South America. Together, these diseases were responsible for over 700 million US dollars in losses in 2001 (4). Commercial cacao production in West Africa and South Asia are not yet affected by WB and FP, but cacao grown in these regions is susceptible to both. With the goal of providing new disease resistant cultivars the USDA-ARS and Mars, Inc. have developed a marker assisted selection (MAS) program. Quantitative trait loci have been identified for resistance to WB, FP, and BP. The potential usefulness of these markers in identifying resistant individuals has been confirmed in an experimental F(1) family in Ecuador.

The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca2+ and Mg2+ dependent-DNase activity and antifungal action on Moniliophthora perniciosa

PubMed Central

2014-01-01

Background The production and accumulation of pathogenesis-related proteins (PR proteins) in plants in response to biotic or abiotic stresses is well known and is considered as a crucial mechanism for plant defense. A pathogenesis-related protein 4 cDNA was identified from a cacao-Moniliophthora perniciosa interaction cDNA library and named TcPR-4b. Results TcPR-4b presents a Barwin domain with six conserved cysteine residues, but lacks the chitin-binding site. Molecular modeling of TcPR-4b confirmed the importance of the cysteine residues to maintain the protein structure, and of several conserved amino acids for the catalytic activity. In the cacao genome, TcPR-4b belonged to a small multigene family organized mainly on chromosome 5. TcPR-4b RT-qPCR analysis in resistant and susceptible cacao plants infected by M. perniciosa showed an increase of expression at 48 hours after infection (hai) in both cacao genotypes. After the initial stage (24-72 hai), the TcPR-4b expression was observed at all times in the resistant genotypes, while in the susceptible one the expression was concentrated at the final stages of infection (45-90 days after infection). The recombinant TcPR-4b protein showed RNase, and bivalent ions dependent-DNase activity, but no chitinase activity. Moreover, TcPR-4b presented antifungal action against M. perniciosa, and the reduction of M. perniciosa survival was related to ROS production in fungal hyphae. Conclusion To our knowledge, this is the first report of a PR-4 showing simultaneously RNase, DNase and antifungal properties, but no chitinase activity. Moreover, we showed that the antifungal activity of TcPR-4b is directly related to RNase function. In cacao, TcPR-4b nuclease activities may be related to the establishment and maintenance of resistance, and to the PCD mechanism, in resistant and susceptible cacao genotypes, respectively. PMID:24920373

Genome-wide analysis reveals divergent patterns of gene expression during zygotic and somatic embryo maturation of Theobroma cacao L., the chocolate tree.

PubMed

Maximova, Siela N; Florez, Sergio; Shen, Xiangling; Niemenak, Nicolas; Zhang, Yufan; Curtis, Wayne; Guiltinan, Mark J

2014-07-16

Theobroma cacao L. is a tropical fruit tree, the seeds of which are used to create chocolate. In vitro somatic embryogenesis (SE) of cacao is a propagation system useful for rapid mass-multiplication to accelerate breeding programs and to provide plants directly to farmers. Two major limitations of cacao SE remain: the efficiency of embryo production is highly genotype dependent and the lack of full cotyledon development results in low embryo to plant conversion rates. With the goal to better understand SE development and to improve the efficiency of SE conversion we examined gene expression differences between zygotic and somatic embryos using a whole genome microarray. The expression of 28,752 genes was determined at 4 developmental time points during zygotic embryogenesis (ZE) and 2 time points during cacao somatic embryogenesis (SE). Within the ZE time course, 10,288 differentially expressed genes were enriched for functions related to responses to abiotic and biotic stimulus, metabolic and cellular processes. A comparison ZE and SE expression profiles identified 10,175 differentially expressed genes. Many TF genes, putatively involved in ethylene metabolism and response, were more strongly expressed in SEs as compared to ZEs. Expression levels of genes involved in fatty acid metabolism, flavonoid biosynthesis and seed storage protein genes were also differentially expressed in the two types of embryos. Large numbers of genes were differentially regulated during various stages of both ZE and SE development in cacao. The relatively higher expression of ethylene and flavonoid related genes during SE suggests that the developing tissues may be experiencing high levels of stress during SE maturation caused by the in vitro environment. The expression of genes involved in the synthesis of auxin, polyunsaturated fatty acids and secondary metabolites was higher in SEs relative to ZEs despite lack of lipid and metabolite accumulation. These differences in gene transcript levels associated with critical processes during seed development are consistent with the fact that somatic embryos do not fully develop the large storage cotyledons found in zygotic embryos. These results provide insight towards design of improved protocols for cacao somatic embryogenesis.

Genome-wide analysis reveals divergent patterns of gene expression during zygotic and somatic embryo maturation of Theobroma cacao L., the chocolate tree

PubMed Central

2014-01-01

Background Theobroma cacao L. is a tropical fruit tree, the seeds of which are used to create chocolate. In vitro somatic embryogenesis (SE) of cacao is a propagation system useful for rapid mass-multiplication to accelerate breeding programs and to provide plants directly to farmers. Two major limitations of cacao SE remain: the efficiency of embryo production is highly genotype dependent and the lack of full cotyledon development results in low embryo to plant conversion rates. With the goal to better understand SE development and to improve the efficiency of SE conversion we examined gene expression differences between zygotic and somatic embryos using a whole genome microarray. Results The expression of 28,752 genes was determined at 4 developmental time points during zygotic embryogenesis (ZE) and 2 time points during cacao somatic embryogenesis (SE). Within the ZE time course, 10,288 differentially expressed genes were enriched for functions related to responses to abiotic and biotic stimulus, metabolic and cellular processes. A comparison ZE and SE expression profiles identified 10,175 differentially expressed genes. Many TF genes, putatively involved in ethylene metabolism and response, were more strongly expressed in SEs as compared to ZEs. Expression levels of genes involved in fatty acid metabolism, flavonoid biosynthesis and seed storage protein genes were also differentially expressed in the two types of embryos. Conclusions Large numbers of genes were differentially regulated during various stages of both ZE and SE development in cacao. The relatively higher expression of ethylene and flavonoid related genes during SE suggests that the developing tissues may be experiencing high levels of stress during SE maturation caused by the in vitro environment. The expression of genes involved in the synthesis of auxin, polyunsaturated fatty acids and secondary metabolites was higher in SEs relative to ZEs despite lack of lipid and metabolite accumulation. These differences in gene transcript levels associated with critical processes during seed development are consistent with the fact that somatic embryos do not fully develop the large storage cotyledons found in zygotic embryos. These results provide insight towards design of improved protocols for cacao somatic embryogenesis. PMID:25030026

Drought tolerance in cacao is mediated by root phenotypic plasticity

USDA-ARS?s Scientific Manuscript database

This study aimed to evaluate phenotypic relationships and their direct and indirect effects through path analysis, and evaluate the use of the phenotypic plasticity index as criteria for the estimation of the basic and explanatory variables used to analysis several cacao progenies subjected to soil ...

Modified pectin from Theobroma cacao induces potent pro-inflammatory activity in murine peritoneal macrophage.

PubMed

Amorim, Juliana C; Vriesmann, Lucia Cristina; Petkowicz, Carmen L O; Martinez, Glaucia Regina; Noleto, Guilhermina R

2016-11-01

In vitro effects of acetylated pectin (OP) isolated from cacao pod husks (Theobroma cacao L.), its partially deacetylated and de-esterified form (MOP), and a commercial homogalacturonan (PG) were investigated on murine peritoneal macrophages. MOP stood out among the studied pectins. After 48h of incubation, compared with the control group, it was able to promote significant macrophage morphological differentiation from resident to activated stage and also stimulated nitric oxide production, which reached a level of 85% of that of LPS stimulus. In the presence of the highest tested concentration of MOP (200μg·mL -1 ), the levels of the cytokines TNF-α (6h) and IL-12 and IL-10 (48h) increased substantially in relation to untreated cells. Our results show that the partial deacetylation and de-esterification of pectin extracted from cacao pod husks (T. cacao L.) produced a polymer with greater ability than its native form to activate macrophages to a cytotoxic phenotype. Like this, they provide the possibility of a therapeutic application to MOP, which could lead to a decreased susceptibility to microbial infection besides antitumor activity. Additionally, the present results also corroborate with the proposition of that the chemical modifications of the biopolymers can result in an improved molecule with new possibilities of application. Copyright © 2016 Elsevier B.V. All rights reserved.

Enhanced resistance in Theobroma cacao against oomycete and fungal pathogens by secretion of phosphatidylinositol-3-phosphate-binding proteins.

PubMed

Helliwell, Emily E; Vega-Arreguín, Julio; Shi, Zi; Bailey, Bryan; Xiao, Shunyuan; Maximova, Siela N; Tyler, Brett M; Guiltinan, Mark J

2016-03-01

The internalization of some oomycete and fungal pathogen effectors into host plant cells has been reported to be blocked by proteins that bind to the effectors' cell entry receptor, phosphatidylinositol-3-phosphate (PI3P). This finding suggested a novel strategy for disease control by engineering plants to secrete PI3P-binding proteins. In this study, we tested this strategy using the chocolate tree Theobroma cacao. Transient expression and secretion of four different PI3P-binding proteins in detached leaves of T. cacao greatly reduced infection by two oomycete pathogens, Phytophthora tropicalis and Phytophthora palmivora, which cause black pod disease. Lesion size and pathogen growth were reduced by up to 85%. Resistance was not conferred by proteins lacking a secretory leader, by proteins with mutations in their PI3P-binding site, or by a secreted PI4P-binding protein. Stably transformed, transgenic T. cacao plants expressing two different PI3P-binding proteins showed substantially enhanced resistance to both P. tropicalis and P. palmivora, as well as to the fungal pathogen Colletotrichum theobromicola. These results demonstrate that secretion of PI3P-binding proteins is an effective way to increase disease resistance in T. cacao, and potentially in other plants, against a broad spectrum of pathogens. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

Novel receptor-like kinases in cacao contain PR-1 extracellular domains.

PubMed

Teixeira, Paulo José Pereira Lima; Costa, Gustavo Gilson Lacerda; Fiorin, Gabriel Lorencini; Pereira, Gonçalo Amarante Guimarães; Mondego, Jorge Maurício Costa

2013-08-01

Members of the pathogenesis-related protein 1 (PR-1) family are well-known markers of plant defence responses, forming part of the arsenal of the secreted proteins produced on pathogen recognition. Here, we report the identification of two cacao (Theobroma cacao L.) PR-1s that are fused to transmembrane regions and serine/threonine kinase domains, in a manner characteristic of receptor-like kinases (RLKs). These proteins (TcPR-1f and TcPR-1g) were named PR-1 receptor kinases (PR-1RKs). Phylogenetic analysis of RLKs and PR-1 proteins from cacao indicated that PR-1RKs originated from a fusion between sequences encoding PR-1 and the kinase domain of a LecRLK (Lectin Receptor-Like Kinase). Retrotransposition marks surround TcPR-1f, suggesting that retrotransposition was involved in the origin of PR-1RKs. Genes with a similar domain architecture to cacao PR-1RKs were found in rice (Oryza sativa), barrel medic (Medicago truncatula) and a nonphototrophic bacterium (Herpetosiphon aurantiacus). However, their kinase domains differed from those found in LecRLKs, indicating the occurrence of convergent evolution. TcPR-1g expression was up-regulated in the biotrophic stage of witches' broom disease, suggesting a role for PR-1RKs during cacao defence responses. We hypothesize that PR-1RKs transduce a defence signal by interacting with a PR-1 ligand. © 2013 BSPP AND JOHN WILEY & SONS LTD.

Moniliophthora perniciosa, the causal agent of witches' broom disease of cacao: what's new from this old foe?

PubMed

Meinhardt, Lyndel W; Rincones, Johana; Bailey, Bryan A; Aime, M Catherine; Griffith, Gareth W; Zhang, Dapeng; Pereira, Gonçalo A G

2008-09-01

Moniliophthora perniciosa (=Crinipellis perniciosa) causes one of the three main fungal diseases of Theobroma cacao (cacao), the source of chocolate. This pathogen causes Witches' broom disease (WBD) and has brought about severe economic losses in all of the cacao-growing regions to which it has spread with yield reductions that range from 50 to 90%. Cacao production in South America reflects the severity of this pathogen, as the yields in most of the infected regions have not returned to pre-outbreak levels, even with the introduction of resistant varieties. In this review we give a brief historical account and summarize the current state of knowledge focusing on developments in the areas of systematics, fungal physiology, biochemistry, genomics and gene expression in an attempt to highlight this disease. Moniliophthora perniciosa is a hemibiotrophic fungus with two distinct growth phases. The ability to culture a biotrophic-like phase in vitro along with new findings derived from the nearly complete genome and expression studies clearly show that these different fungal growth phases function under distinct metabolic parameters. These new findings have greatly improved our understanding of this fungal/host interaction and we may be at the crossroads of understanding how hemibiotrophic fungal plant pathogens cause disease in other crops. The first WDB symptoms appear to have been described in the diaries of Alexandre Rodrigues Ferreira (described as lagartão; meaning big lizard) from his observations of cacao trees in 1785 and 1787 in Amazonia, which is consistent with the generally accepted idea that M. perniciosa, like its main host T. cacao, evolved in this region. The disease subsequently arrived in Surinam in 1895. WBD moved rapidly, spreading to Guyana in 1906, Ecuador in 1918, Trinidad in 1928, Colombia in 1929 and Grenada in 1948. In each case, cacao production was catastrophically affected with yield reductions of 50-90%. After the arrival of M. perniciosa in Bahia in 1989, Brazil went from being the world's 3rd largest producer of cacao (347 000 tonnes in 1988-1990; c. 15% of the total world production at that time) to a net importer (141 000 tonnes in 1998-2000). Fortunately for chocolate lovers, other regions of the world such as West Africa and South East Asia have not yet been affected by this disease and have expanded production to meet growing world demand (predicted to reach 3 700 000 tonnes by 2010). Moniliophthora perniciosa (Stahel) Aime & Phillips-Mora: super-kingdom Eukaryota; kingdom Fungi; phylum Basidiomycota; subphylum Agaricomycotina; class Agaricomycetes; subclass Agaricomycetidae; order Agaricales; family Marasmiaceae; genus Moniliophthora. http://www.lge.ibi.unicamp.br/vassoura/, http://nt.ars-grin.gov/taxadescriptions/keys/TrichodermaIndex.cfm, http://www.worldcocoafoundation.org/info-center/research-updates.asp, http://www.ars.usda.gov/ba/psi/spcl.

Theobroma cacao: A genetically integrated physical map and genome-scale comparative synteny analysis

USDA-ARS?s Scientific Manuscript database

A comprehensive integrated genomic framework is considered a centerpiece of genomic research. In collaboration with the USDA-ARS (SHRS) and Mars Inc., the Clemson University Genomics Institute (CUGI) has developed a genetically anchored physical map of the T. cacao genome. Three BAC libraries contai...

21 CFR 163.111 - Chocolate liquor.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Chocolate liquor. 163.111 Section 163.111 Food and... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.111 Chocolate liquor. (a) Description. (1) Chocolate liquor is the solid or semiplastic food prepared by finely grinding...

21 CFR 163.111 - Chocolate liquor.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Chocolate liquor. 163.111 Section 163.111 Food and... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.111 Chocolate liquor. (a) Description. (1) Chocolate liquor is the solid or semiplastic food prepared by finely grinding...

21 CFR 163.111 - Chocolate liquor.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Chocolate liquor. 163.111 Section 163.111 Food and... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.111 Chocolate liquor. (a) Description. (1) Chocolate liquor is the solid or semiplastic food prepared by finely grinding...

Characterization of naturalized cacao populations in Puerto Rico

USDA-ARS?s Scientific Manuscript database

Native to the headwaters of the Amazon River, cacao is an important agricultural tree crop produced in tropical regions around the world. Its raw product, the seed or ‘beans’, is the source for the multi-billion dollar chocolate industry. The USDA-ARS Tropical Agriculture Research Station in Mayag...

The influence of formulation on Trichoderma biological activity and frosty pod rot disease management in Theobroma cacao

USDA-ARS?s Scientific Manuscript database

Frosty pod rot (FPR), caused by Moniliophthora roreri is responsible for significant losses in Theobroma cacao. Due to the limited options for FPR management, biological control methods using Trichoderma are being studied. Combinations of three formulations and two Trichoderma isolates were studied ...

40 CFR 180.491 - Propylene oxide; tolerances for residues.

Code of Federal Regulations, 2011 CFR

2011-07-01

...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances... commodities: Commodity Parts per million Basil, dried leaves 6000 Cacao bean, dried bean 20.0 Cacao bean... basil 1500 Nutmeat, processed, except peanuts 10.0 Onion, dried 6000 Plum, prune, dried 2.0 (b) Section...

Soil classification and carbon storage in cacao agroforestry farming systems of Bahia, Brazil

USDA-ARS?s Scientific Manuscript database

Information concerning the classification of soils and their properties under cacao agroforestry systems of the Atlantic rain forest biome region in the Southeast of Bahia Brazil is largely unknown. Soil and climatic conditions in this region are favorable for high soil carbon storage. This study is...

Register of new fruit and nut cultivars list 48. Banana, cacao, plantain

USDA-ARS?s Scientific Manuscript database

The Register of New Fruit and Nut Varieties 48 is a compilation of descriptions of new fruit and nut cultivars from around the world. In this edition, newly released banana, plantain, and cacao cultivars are described in terms of their origins, important fruit traits and yield. ...

Increasing the efficiency of traditional cacao breeding using whole genome sequencing information.

USDA-ARS?s Scientific Manuscript database

Unfortunately, 80% of the genotypes from the "hybrid seeds" are unproductive in farmers' fields. In 1999 the USDA and Mars Inc. initiated a Marker Assisted Selection (MAS) program for cacao that has reduces many of the problems in traditional breeding. One limitation of the MAS program is the distan...

Identification of candidate genes involved in witches’ broom disease resistance in a segregating mapping population of Theobroma cacao L. in Brazil

USDA-ARS?s Scientific Manuscript database

Witches’ broom disease (WBD) caused by the fungus Moniliophthora perniciosa is responsible for considerable economic losses for cacao producers in the Americas. Protective fungicides are ineffective, and disease management involving repeated phytosanitary removals increases labor costs. The best al...

Phytophthora megakarya, a causal agent of black pod rot in Africa

USDA-ARS?s Scientific Manuscript database

In most parts of the world where Theobroma cacao is grown, Phytophthora palmivora is the major concern for causing black pod rot (BPR). Phytophthora megakarya, on the other hand, occurs only in Africa, but represents a major threat to cacao production, the countries of West Africa being the largest ...

Microsatellite-aided detection of genetic redundancy improves management of the International Cocoa Genebank, Trinidad

USDA-ARS?s Scientific Manuscript database

Cacao (Theobroma cacao L.), the tree from which cocoa butter and chocolate is derived, is conserved in field genebanks. The largest of these ex situ collections in the public domain is the International Cocoa Genebank, Trinidad (ICG,T). Reduction of genetic redundancy is essential to improve the acc...

Applying SNP marker technology in the cacao breeding program at the Cocoa Research Institute of Ghana

USDA-ARS?s Scientific Manuscript database

In this investigation 45 parental cacao plants and five progeny derived from the parental stock studied were genotyped using six SNP markers to determine off-types or mislabeled clones and to authenticate crosses made in the Cocoa Research Institute of Ghana (CRIG) breeding program. Investigation wa...

21 CFR 163.150 - Sweet cocoa and vegetable fat coating.

Code of Federal Regulations, 2010 CFR

2010-04-01

... preparation of the product, cocoa or a mixture of cocoa and chocolate liquor is used in such quantity that the... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Sweet cocoa and vegetable fat coating. 163.150... (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products...

21 CFR 163.113 - Cocoa.

Code of Federal Regulations, 2010 CFR

2010-04-01

... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.113 Cocoa. (a) Description. Cocoa is the food that conforms to the definition and standard of identity, and is subject to the... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Cocoa. 163.113 Section 163.113 Food and Drugs FOOD...

21 CFR 163.114 - Lowfat cocoa.

Code of Federal Regulations, 2010 CFR

2010-04-01

... CONSUMPTION CACAO PRODUCTS Requirements for Specific Standardized Cacao Products § 163.114 Lowfat cocoa. (a) Description. Lowfat cocoa is the food that conforms to the definition and standard of identity, and is subject... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Lowfat cocoa. 163.114 Section 163.114 Food and...

The impact of SNP fingerprinting and parentage analysis on the effectiveness of variety recommendations in cacao

USDA-ARS?s Scientific Manuscript database

Evidence for the impact of mislabeling and/or pollen contamination on consistency of field performance has been lacking to reinforce the need for strict adherence to quality control protocols in cacao seed garden and germplasm plot management. The present study used SNP fingerprinting at 64 loci to ...

40 CFR 180.491 - Propylene oxide; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances... Basil, dried leaves 6000 Cacao bean, cocoa powder 20.0 Cacao bean, dried bean 20.0 Fig 3.0 Garlic, dried 6000 Grape, raisin 4.0 Herbs and spices, group 19, dried, except basil 1500 Nut, pine 10.0 Nut, tree...

40 CFR 180.491 - Propylene oxide; tolerances for residues.

Code of Federal Regulations, 2010 CFR

2010-07-01

...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances...: Commodity Parts per million Basil, dried leaves 6000 Cacao bean, dried bean 20.0 Cacao bean, cocoa powder 20.0 Fig 3.0 Garlic, dried 6000 Grape, raisin 4.0 Herbs and spices, group 19, dried, except basil 1500...

40 CFR 180.491 - Propylene oxide; tolerances for residues.

Code of Federal Regulations, 2012 CFR

2012-07-01

...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances... Basil, dried leaves 6000 Cacao bean, cocoa powder 20.0 Cacao bean, dried bean 20.0 Fig 3.0 Garlic, dried 6000 Grape, raisin 4.0 Herbs and spices, group 19, dried, except basil 1500 Nut, pine 10.0 Nut, tree...

40 CFR 180.491 - Propylene oxide; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances... Basil, dried leaves 6000 Cacao bean, cocoa powder 20.0 Cacao bean, dried bean 20.0 Fig 3.0 Garlic, dried 6000 Grape, raisin 4.0 Herbs and spices, group 19, dried, except basil 1500 Nut, pine 10.0 Nut, tree...

Toward The identification Of candidate genes involved in black pod disease resistance in Theobroma cacao L.

USDA-ARS?s Scientific Manuscript database

Increasing yield, quality and disease resistance are important objectives for cacao breeding programs. Some of the diseases, such as black pod rot (Phytophtora spp), frosty pod (Moniliophthora roreri) and witches’ broom (M. perniciosa), produce significant losses in all or in some of the various pro...

Morphological variants of Moniliophthora roreri on artificial media and the biotroph/necrotroph shift

USDA-ARS?s Scientific Manuscript database

Moniliophthora roreri is an unusual fungus and plant pathogen on many levels. Although M. roreri is an aggressive pathogen of cacao (Theobroma cacao), many species grouped in the genus Moniliophthora are considered endophytes. The morphology of M. roreri is confusing and for many years it was consid...

21 CFR 163.5 - Methods of analysis.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Methods of analysis. 163.5 Section 163.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS General Provisions § 163.5 Methods of analysis. Shell and cacao fat content in...

21 CFR 163.5 - Methods of analysis.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Methods of analysis. 163.5 Section 163.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS General Provisions § 163.5 Methods of analysis. Shell and cacao fat content in...

21 CFR 163.5 - Methods of analysis.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Methods of analysis. 163.5 Section 163.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS General Provisions § 163.5 Methods of analysis. Shell and cacao fat content in...

21 CFR 163.5 - Methods of analysis.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Methods of analysis. 163.5 Section 163.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS General Provisions § 163.5 Methods of analysis. Shell and cacao fat content in...

Ultra-barcoding in cacao (Theobroma spp.; malvaceae) using whole chloroplast genomes and nuclear ribosomal DNA

USDA-ARS?s Scientific Manuscript database

High-throughput next-generation sequencing was used to scan the genome and generate reliable sequence of high copy number regions. Using this method, we examined whole plastid genomes as well as nearly 6000 bases of nuclear ribosomal DNA sequences for nine genotypes of Theobroma cacao and an indivi...

Register of New Fruit and Nut Cultivars List 45. Banana, cacao, Spanish lime, plantain

USDA-ARS?s Scientific Manuscript database

The Register of New Fruit and Nut Varieties 45 is a compilation of descriptions of new fruit and nut cultivars from around the world. In this edition, newly released cacao, banana, plantain, and genip cultivars are described in terms of their origins, important fruit traits and yield....

Genetic identity and parentage in farmer selections of cacao from Southern Sulawesi, Indonesia revealed by molecular markers

USDA-ARS?s Scientific Manuscript database

Indonesia is the 3rd largest cocoa producing countries in the world and 71% of its production is from Sulawesi Island. Knowledge about the genetic background of farmer selections is highly important for effective identification and rational deployment of superior cacao clones in farmers’ fields. Mor...

How dark chocolate is processed

USDA-ARS?s Scientific Manuscript database

This month’s column will continue the theme of “How Is It Processed?” The column will focus on dark chocolate. The botanical name for the cacao tree is Theobroma cacao, which literally means “food of the Gods.” Dark chocolate is both delicious and nutritious. Production of dark chocolate will be des...

The sorption and desorption of phosphate-P, ammonium-N and nitrate-N in cacao shell and corn cob biochars.

PubMed

Hale, S E; Alling, V; Martinsen, V; Mulder, J; Breedveld, G D; Cornelissen, G

2013-06-01

The sorption of PO4-P, NH4-N and NO3-N to cacao shell and corn cob biochars produced at 300-350°C was quantified. The biochars were used; (i) as received (unwashed), (ii) after rinsing with Millipore water and (iii) following leaching with Millipore water. In addition to sorption, desorption of PO4-P from the unwashed biochars was quantified. There was no sorption of PO4-P to either washed or rinsed biochars, but following leaching, both biochars adsorbed PO4-P and distribution coefficients (Kd L kg(-1)) were very similar for both materials (10(1.1±0.5) for cacao shell biochar and 10(1.0±0.2) for corn cob biochar). The BET surface area and micropore volume increased 80% and 60% for the cacao shell and corn cob biochars following leaching. After 60 d, 1483±45 mg kg(-1) and 172±1 mg kg(-1) PO4-P was released from the cacao shell and corn cob biochars. NH4-N was sorbed by both unwashed biochars, albeit weakly with Kd values around 10(2) L kg(-1). We speculate that NH4-N could bind via an electrostatic exchange with other cationic species on the surface of the biochar. There was no significant release or sorption of NO3-N from or to either of the biochars. Copyright © 2013 Elsevier Ltd. All rights reserved.

Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds

PubMed Central

Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

2016-01-01

Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents. PMID:26947442

Impact of fermentation, drying, roasting, and Dutch processing on epicatechin and catechin content of cacao beans and cocoa ingredients.

PubMed

Payne, Mark J; Hurst, W Jeffrey; Miller, Kenneth B; Rank, Craig; Stuart, David A

2010-10-13

Low molecular weight flavan-3-ols are thought to be responsible, in part, for the cardiovascular benefits associated with cocoa powder and dark chocolate. The levels of epicatechin and catechin were determined in raw and conventionally fermented cacao beans and during conventional processing, which included drying, roasting, and Dutch (alkali) processing. Unripe cacao beans had 29% higher levels of epicatechin and the same level of catechin compared to fully ripe beans. Drying had minimal effect on the epicatechin and catechin levels. Substantial decreases (>80%) in catechin and epicatechin levels were observed in fermented versus unfermented beans. When both Ivory Coast and Papua New Guinea beans were subjected to roasting under controlled conditions, there was a distinct loss of epicatechin when bean temperatures exceeded 70 °C. When cacao beans were roasted to 120 °C, the catechin level in beans increased by 696% in unfermented beans, by 650% in Ivory Coast beans, and by 640% in Papua New Guinea fermented beans compared to the same unroasted beans. These results suggest that roasting in excess of 70 °C generates significant amounts of (-)-catechin, probably due to epimerization of (-)-epicatechin. Compared to natural cocoa powders, Dutch processing caused a loss in both epicatechin (up to 98%) and catechin (up to 80%). The epicatechin/catechin ratio is proposed as a useful and sensitive indicator for the processing history of cacao beans.

Ultra-barcoding in cacao (Theobroma spp.; Malvaceae) using whole chloroplast genomes and nuclear ribosomal DNA.

PubMed

Kane, Nolan; Sveinsson, Saemundur; Dempewolf, Hannes; Yang, Ji Yong; Zhang, Dapeng; Engels, Johannes M M; Cronk, Quentin

2012-02-01

To reliably identify lineages below the species level such as subspecies or varieties, we propose an extension to DNA-barcoding using next-generation sequencing to produce whole organellar genomes and substantial nuclear ribosomal sequence. Because this method uses much longer versions of the traditional DNA-barcoding loci in the plastid and ribosomal DNA, we call our approach ultra-barcoding (UBC). We used high-throughput next-generation sequencing to scan the genome and generate reliable sequence of high copy number regions. Using this method, we examined whole plastid genomes as well as nearly 6000 bases of nuclear ribosomal DNA sequences for nine genotypes of Theobroma cacao and an individual of the related species T. grandiflorum, as well as an additional publicly available whole plastid genome of T. cacao. All individuals of T. cacao examined were uniquely distinguished, and evidence of reticulation and gene flow was observed. Sequence variation was observed in some of the canonical barcoding regions between species, but other regions of the chloroplast were more variable both within species and between species, as were ribosomal spacers. Furthermore, no single region provides the level of data available using the complete plastid genome and rDNA. Our data demonstrate that UBC is a viable, increasingly cost-effective approach for reliably distinguishing varieties and even individual genotypes of T. cacao. This approach shows great promise for applications where very closely related or interbreeding taxa must be distinguished.

Morphological, Physiological, and Taxonomic Characterization of Actinobacterial Isolates Living as Endophytes of Cacao Pods and Cacao Seeds.

PubMed

Tchinda, Romaric Armel Mouafo; Boudjeko, Thaddée; Simao-Beaunoir, Anne-Marie; Lerat, Sylvain; Tsala, Éric; Monga, Ernest; Beaulieu, Carole

2016-01-01

Vascular plants are commonly colonized by endophytic actinobacteria. However, very little is known about the relationship between these microorganisms and cacao fruits. In order to determine the physiological and taxonomic relationships between the members of this community, actinobacteria were isolated from cacao fruits and seeds. Among the 49 isolates recovered, 11 morphologically distinct isolates were selected for further characterization. Sequencing of the 16S rRNA gene allowed the partition of the selected isolates into three phylogenetic clades. Most of the selected endophytic isolates belonged to the Streptomyces violaceusniger clade. Physiological characterization was carried out and a similarity index was used to cluster the isolates. However, clustering based on physiological properties did not match phylogenetic lineages. Isolates were also characterized for traits commonly associated with plant growth-promoting bacteria, including antibiosis and auxin biosynthesis. All isolates exhibited resistance to geldanamycin, whereas only two isolates were shown to produce this antibiotic. Endophytes were inoculated on radish seedlings and most isolates were found to possess plant growth-promoting abilities. These endophytic actinobacteria inhibited the growth of various plant pathogenic fungi and/or bacteria. The present study showed that S. violaceusniger clade members represent a significant part of the actinobacterial community living as endophytes in cacao fruits and seeds. While several members of this clade are known to be geldanamycin producers and efficient biocontrol agents of plant diseases, we herein established the endophytic lifestyle of some of these microorganisms, demonstrating their potential as plant health agents.

Tree spatial structure, host composition and resource availability influence mirid density or black pod prevalence in cacao agroforests in Cameroon.

PubMed

Gidoin, Cynthia; Babin, Régis; Bagny Beilhe, Leïla; Cilas, Christian; ten Hoopen, Gerben Martijn; Bieng, Marie Ange Ngo

2014-01-01

Combining crop plants with other plant species in agro-ecosystems is one way to enhance ecological pest and disease regulation mechanisms. Resource availability and microclimatic variation mechanisms affect processes related to pest and pathogen life cycles. These mechanisms are supported both by empirical research and by epidemiological models, yet their relative importance in a real complex agro-ecosystem is still not known. Our aim was thus to assess the independent effects and the relative importance of different variables related to resource availability and microclimatic variation that explain pest and disease occurrence at the plot scale in real complex agro-ecosystems. The study was conducted in cacao (Theobroma cacao) agroforests in Cameroon, where cocoa production is mainly impacted by the mirid bug, Sahlbergella singularis, and black pod disease, caused by Phytophthora megakarya. Vegetation composition and spatial structure, resource availability and pest and disease occurrence were characterized in 20 real agroforest plots. Hierarchical partitioning was used to identify the causal variables that explain mirid density and black pod prevalence. The results of this study show that cacao agroforests can be differentiated on the basis of vegetation composition and spatial structure. This original approach revealed that mirid density decreased when a minimum number of randomly distributed forest trees were present compared with the aggregated distribution of forest trees, or when forest tree density was low. Moreover, a decrease in mirid density was also related to decreased availability of sensitive tissue, independently of the effect of forest tree structure. Contrary to expectations, black pod prevalence decreased with increasing cacao tree abundance. By revealing the effects of vegetation composition and spatial structure on mirids and black pod, this study opens new perspectives for the joint agro-ecological management of cacao pests and diseases at the plot scale, through the optimization of the spatial structure and composition of the vegetation.

Prediction of Cacao (Theobroma cacao) Resistance to Moniliophthora spp. Diseases via Genome-Wide Association Analysis and Genomic Selection.

PubMed

McElroy, Michel S; Navarro, Alberto J R; Mustiga, Guiliana; Stack, Conrad; Gezan, Salvador; Peña, Geover; Sarabia, Widem; Saquicela, Diego; Sotomayor, Ignacio; Douglas, Gavin M; Migicovsky, Zoë; Amores, Freddy; Tarqui, Omar; Myles, Sean; Motamayor, Juan C

2018-01-01

Cacao ( Theobroma cacao ) is a globally important crop, and its yield is severely restricted by disease. Two of the most damaging diseases, witches' broom disease (WBD) and frosty pod rot disease (FPRD), are caused by a pair of related fungi: Moniliophthora perniciosa and Moniliophthora roreri , respectively. Resistant cultivars are the most effective long-term strategy to address Moniliophthora diseases, but efficiently generating resistant and productive new cultivars will require robust methods for screening germplasm before field testing. Marker-assisted selection (MAS) and genomic selection (GS) provide two potential avenues for predicting the performance of new genotypes, potentially increasing the selection gain per unit time. To test the effectiveness of these two approaches, we performed a genome-wide association study (GWAS) and GS on three related populations of cacao in Ecuador genotyped with a 15K single nucleotide polymorphism (SNP) microarray for three measures of WBD infection (vegetative broom, cushion broom, and chirimoya pod), one of FPRD (monilia pod) and two productivity traits (total fresh weight of pods and % healthy pods produced). GWAS yielded several SNPs associated with disease resistance in each population, but none were significantly correlated with the same trait in other populations. Genomic selection, using one population as a training set to estimate the phenotypes of the remaining two (composed of different families), varied among traits, from a mean prediction accuracy of 0.46 (vegetative broom) to 0.15 (monilia pod), and varied between training populations. Simulations demonstrated that selecting seedlings using GWAS markers alone generates no improvement over selecting at random, but that GS improves the selection process significantly. Our results suggest that the GWAS markers discovered here are not sufficiently predictive across diverse germplasm to be useful for MAS, but that using all markers in a GS framework holds substantial promise in accelerating disease-resistance in cacao.

Tc-MYBPA an Arabidopsis TT2-like transcription factor and functions in the regulation of proanthocyanidin synthesis in Theobroma cacao.

PubMed

Liu, Yi; Shi, Zi; Maximova, Siela N; Payne, Mark J; Guiltinan, Mark J

2015-06-25

The flavan-3-ols catechin and epicatechin, and their polymerized oligomers, the proanthocyanidins (PAs, also called condensed tannins), accumulate to levels of up to 15 % of the total weight of dry seeds of Theobroma cacao L. These compounds have been associated with several health benefits in humans. They also play important roles in pest and disease defense throughout the plant. In Arabidopsis, the R2R3 type MYB transcription factor TT2 regulates the major genes leading to the synthesis of PA. To explore the transcriptional regulation of the PA synthesis pathway in cacao, we isolated and characterized an R2R3 type MYB transcription factor MYBPA from cacao. We examined the spatial and temporal gene expression patterns of the Tc-MYBPA gene and found it to be developmentally expressed in a manner consistent with its involvement in PAs and anthocyanin synthesis. Functional complementation of an Arabidopsis tt2 mutant with Tc-MYBPA suggested that it can functionally substitute the Arabidopsis TT2 gene. Interestingly, in addition to PA accumulation in seeds of the Tc-MYBPA expressing plants, we also observed an obvious increase of anthocyanidin accumulation in hypocotyls. We observed that overexpression of the Tc-MYBPA gene resulted in increased expression of several key genes encoding the major structural enzymes of the PA and anthocyanidin pathway, including DFR (dihydroflavanol reductase), LDOX (leucoanthocyanidin dioxygenase) and BAN (ANR, anthocyanidin reductase). We conclude that the Tc-MYBPA gene that encodes an R2R3 type MYB transcription factor is an Arabidopsis TT2 like transcription factor, and may be involved in the regulation of both anthocyanin and PA synthesis in cacao. This research may provide molecular tools for breeding of cacao varieties with improved disease resistance and enhanced flavonoid profiles for nutritional and pharmaceutical applications.

Enhanced somatic embryogenesis in Theobroma cacao using the homologous BABY BOOM transcription factor.

PubMed

Florez, Sergio L; Erwin, Rachel L; Maximova, Siela N; Guiltinan, Mark J; Curtis, Wayne R

2015-05-16

Theobroma cacao, the chocolate tree, is an important economic crop in East Africa, South East Asia, and South and Central America. Propagation of elite varieties has been achieved through somatic embryogenesis (SE) but low efficiencies and genotype dependence still presents a significant limitation for its propagation at commercial scales. Manipulation of transcription factors has been used to enhance the formation of SEs in several other plant species. This work describes the use of the transcription factor Baby Boom (BBM) to promote the transition of somatic cacao cells from the vegetative to embryonic state. An ortholog of the Arabidopsis thaliana BBM gene (AtBBM) was characterized in T. cacao (TcBBM). TcBBM expression was observed throughout embryo development and was expressed at higher levels during SE as compared to zygotic embryogenesis (ZE). TcBBM overexpression in A. thaliana and T. cacao led to phenotypes associated with SE that did not require exogenous hormones. While transient ectopic expression of TcBBM provided only moderate enhancements in embryogenic potential, constitutive overexpression dramatically increased SE proliferation but also appeared to inhibit subsequent development. Our work provides validation that TcBBM is an ortholog to AtBBM and has a specific role in both somatic and zygotic embryogenesis. Furthermore, our studies revealed that TcBBM transcript levels could serve as a biomarker for embryogenesis in cacao tissue. Results from transient expression of TcBBM provide confirmation that transcription factors can be used to enhance SE without compromising plant development and avoiding GMO plant production. This strategy could compliment a hormone-based method of reprogramming somatic cells and lead to more precise manipulation of SE at the regulatory level of transcription factors. The technology would benefit the propagation of elite varieties with low regeneration potential as well as the production of transgenic plants, which similarly requires somatic cell reprogramming.

TcNPR3 from Theobroma cacao functions as a repressor of the pathogen defense response

PubMed Central

2013-01-01

Background Arabidopsis thaliana (Arabidopsis) NON-EXPRESSOR OF PR1 (NPR1) is a transcription coactivator that plays a central role in regulating the transcriptional response to plant pathogens. Developing flowers of homozygous npr3 mutants are dramatically more resistant to infection by the pathogenic bacterium Pseudomonas syringae, suggesting a role of NPR3 as a repressor of NPR1-mediated defense response with a novel role in flower development. Results We report here the characterization of a putative NPR3 gene from the tropical tree species Theobroma cacao (TcNPR3). Like in Arabidopsis, TcNPR3 was constitutively expressed across a wide range of tissue types and developmental stages but with some differences in relative levels compared to Arabidopsis. To test the function of TcNPR3, we performed transgenic complementation analysis by introducing a constitutively expressing putative TcNPR3 transgene into an Arabidopsis npr3 mutant. TcNPR3 expressing Arabidopsis plants were partially restored to the WT pathogen phenotype (immature flowers susceptible to bacterial infection). To test TcNPR3 function directly in cacao tissues, a synthetic microRNA targeting TcNPR3 mRNA was transiently expressed in cacao leaves using an Agrobacterium-infiltration method. TcNPR3 knock down leaf tissues were dramatically more resistance to infection with Phytophthora capsici in a leaf bioassay, showing smaller lesion sizes and reduced pathogen replication. Conclusions We conclude that TcNPR3 functions similar to the Arabidopsis NPR3 gene in the regulation of the cacao defense response. Since TcNPR3 did not show a perfect complementation of the Arabidopsis NPR3 mutation, the possibility remains that other functions of TcNPR3 remain to be found. This novel knowledge can contribute to the breeding of resistant cacao varieties against pathogens through molecular markers based approaches or biotechnological strategies. PMID:24314063

Tree Spatial Structure, Host Composition and Resource Availability Influence Mirid Density or Black Pod Prevalence in Cacao Agroforests in Cameroon

PubMed Central

Gidoin, Cynthia; Babin, Régis; Bagny Beilhe, Leïla; Cilas, Christian; ten Hoopen, Gerben Martijn; Bieng, Marie Ange Ngo

2014-01-01

Combining crop plants with other plant species in agro-ecosystems is one way to enhance ecological pest and disease regulation mechanisms. Resource availability and microclimatic variation mechanisms affect processes related to pest and pathogen life cycles. These mechanisms are supported both by empirical research and by epidemiological models, yet their relative importance in a real complex agro-ecosystem is still not known. Our aim was thus to assess the independent effects and the relative importance of different variables related to resource availability and microclimatic variation that explain pest and disease occurrence at the plot scale in real complex agro-ecosystems. The study was conducted in cacao (Theobroma cacao) agroforests in Cameroon, where cocoa production is mainly impacted by the mirid bug, Sahlbergella singularis, and black pod disease, caused by Phytophthora megakarya. Vegetation composition and spatial structure, resource availability and pest and disease occurrence were characterized in 20 real agroforest plots. Hierarchical partitioning was used to identify the causal variables that explain mirid density and black pod prevalence. The results of this study show that cacao agroforests can be differentiated on the basis of vegetation composition and spatial structure. This original approach revealed that mirid density decreased when a minimum number of randomly distributed forest trees were present compared with the aggregated distribution of forest trees, or when forest tree density was low. Moreover, a decrease in mirid density was also related to decreased availability of sensitive tissue, independently of the effect of forest tree structure. Contrary to expectations, black pod prevalence decreased with increasing cacao tree abundance. By revealing the effects of vegetation composition and spatial structure on mirids and black pod, this study opens new perspectives for the joint agro-ecological management of cacao pests and diseases at the plot scale, through the optimization of the spatial structure and composition of the vegetation. PMID:25313514

TcNPR3 from Theobroma cacao functions as a repressor of the pathogen defense response.

PubMed

Shi, Zi; Zhang, Yufan; Maximova, Siela N; Guiltinan, Mark J

2013-12-06

Arabidopsis thaliana (Arabidopsis) NON-EXPRESSOR OF PR1 (NPR1) is a transcription coactivator that plays a central role in regulating the transcriptional response to plant pathogens. Developing flowers of homozygous npr3 mutants are dramatically more resistant to infection by the pathogenic bacterium Pseudomonas syringae, suggesting a role of NPR3 as a repressor of NPR1-mediated defense response with a novel role in flower development. We report here the characterization of a putative NPR3 gene from the tropical tree species Theobroma cacao (TcNPR3). Like in Arabidopsis, TcNPR3 was constitutively expressed across a wide range of tissue types and developmental stages but with some differences in relative levels compared to Arabidopsis. To test the function of TcNPR3, we performed transgenic complementation analysis by introducing a constitutively expressing putative TcNPR3 transgene into an Arabidopsis npr3 mutant. TcNPR3 expressing Arabidopsis plants were partially restored to the WT pathogen phenotype (immature flowers susceptible to bacterial infection). To test TcNPR3 function directly in cacao tissues, a synthetic microRNA targeting TcNPR3 mRNA was transiently expressed in cacao leaves using an Agrobacterium-infiltration method. TcNPR3 knock down leaf tissues were dramatically more resistance to infection with Phytophthora capsici in a leaf bioassay, showing smaller lesion sizes and reduced pathogen replication. We conclude that TcNPR3 functions similar to the Arabidopsis NPR3 gene in the regulation of the cacao defense response. Since TcNPR3 did not show a perfect complementation of the Arabidopsis NPR3 mutation, the possibility remains that other functions of TcNPR3 remain to be found. This novel knowledge can contribute to the breeding of resistant cacao varieties against pathogens through molecular markers based approaches or biotechnological strategies.

The Theobroma cacao B3 domain transcription factor TcLEC2 plays a duel role in control of embryo development and maturation

PubMed Central

2014-01-01

Background The Arabidopsis thaliana LEC2 gene encodes a B3 domain transcription factor, which plays critical roles during both zygotic and somatic embryogenesis. LEC2 exerts significant impacts on determining embryogenic potential and various metabolic processes through a complicated genetic regulatory network. Results An ortholog of the Arabidopsis Leafy Cotyledon 2 gene (AtLEC2) was characterized in Theobroma cacao (TcLEC2). TcLEC2 encodes a B3 domain transcription factor preferentially expressed during early and late zygotic embryo development. The expression of TcLEC2 was higher in dedifferentiated cells competent for somatic embryogenesis (embryogenic calli), compared to non-embryogenic calli. Transient overexpression of TcLEC2 in immature zygotic embryos resulted in changes in gene expression profiles and fatty acid composition. Ectopic expression of TcLEC2 in cacao leaves changed the expression levels of several seed related genes. The overexpression of TcLEC2 in cacao explants greatly increased the frequency of regeneration of stably transformed somatic embryos. TcLEC2 overexpressing cotyledon explants exhibited a very high level of embryogenic competency and when cultured on hormone free medium, exhibited an iterative embryogenic chain-reaction. Conclusions Our study revealed essential roles of TcLEC2 during both zygotic and somatic embryo development. Collectively, our evidence supports the conclusion that TcLEC2 is a functional ortholog of AtLEC2 and that it is involved in similar genetic regulatory networks during cacao somatic embryogenesis. To our knowledge, this is the first detailed report of the functional analysis of a LEC2 ortholog in a species other then Arabidopsis. TcLEC2 could potentially be used as a biomarker for the improvement of the SE process and screen for elite varieties in cacao germplasm. PMID:24758406

Molecular, physiological and morphological analysis of waterlogging tolerance in clonal genotypes of Theobroma cacao L.

PubMed

Bertolde, Fabiana Zanelato; De Almeida, Alex-Alan Furtado; Corrêa, Ronan Xavier; Gomes, Fábio Pinto; Gaiotto, Fernanda Amato; Baligar, Virupax C; Loguercio, Leandro Lopes

2010-01-01

In soil, anoxia conditions generated by waterlogging induce changes in genetic, morphological and physiological processes, altering the growth and development of plants. Mass propagation of cacao (Theobroma cacao L.) plantlets (clones) is affected by waterlogging caused by heavy rains and irrigation methods used to induce rooting. An experiment was undertaken to assess the effects of a 45-day flooding (anoxia) on physiological and morphological traits of 35 elite cacao genotypes, aiming at potentially identifying those with greater tolerance to flooding of the growth substrate. Eighteen fluorochrome-labeled microsatellite (SSR) primer pairs were used to assess genetic variability among clones, with 248 alleles being amplified and used to calculate similarity coefficients. The resulting dendrogram indicated the presence of four major groups, in which two represented 60% and 31% of the genotypes tested. A general trend toward high levels of heterozygosity was also found for physiological and morphological traits. The survival index (IS) for flood tolerance observed varied from 30 to 96%. Clones TSA-654, TSA-656, TSA-792, CA-1.4, CEPEC-2009 and PH-17 showed an IS value above 94%, whereas CEPEC-2010, CEPEC-2002, CA-7.1 and VB-903 clones were those mostly affected by waterlogging, with IS value below 56%. All genotypes displayed lenticel and adventitious root formation in response to waterlogging, although with different intensities. To determine whether patterns of physiological response could be associated with tolerance to anoxia, a similarity-grouping analysis was performed using the ratio between waterlogged and control values obtained for a series of physiological variables assessed. No specific pattern of physiological and morphological responses to waterlogging was strictly associated with survival of plantlets. However, results revealed by the dendrogram suggest that absence of leaf chlorosis may be a proper trait to indicate cacao clones with higher survival rates under flooding conditions. Consequences of these findings are discussed in the context of developing improved strategies for mass production of clones from elite cacao genotypes.

Functional analysis of the theobroma cacao NPR1 gene in arabidopsis

PubMed Central

2010-01-01

Background The Arabidopsis thaliana NPR1 gene encodes a transcription coactivator (NPR1) that plays a major role in the mechanisms regulating plant defense response. After pathogen infection and in response to salicylic acid (SA) accumulation, NPR1 translocates from the cytoplasm into the nucleus where it interacts with other transcription factors resulting in increased expression of over 2000 plant defense genes contributing to a pathogen resistance response. Results A putative Theobroma cacao NPR1 cDNA was isolated by RT-PCR using degenerate primers based on homologous sequences from Brassica, Arabidopsis and Carica papaya. The cDNA was used to isolate a genomic clone from Theobroma cacao containing a putative TcNPR1 gene. DNA sequencing revealed the presence of a 4.5 kb coding region containing three introns and encoding a polypeptide of 591 amino acids. The predicted TcNPR1 protein shares 55% identity and 78% similarity to Arabidopsis NPR1, and contains each of the highly conserved functional domains indicative of this class of transcription factors (BTB/POZ and ankyrin repeat protein-protein interaction domains and a nuclear localization sequence (NLS)). To functionally define the TcNPR1 gene, we transferred TcNPR1 into an Arabidopsis npr1 mutant that is highly susceptible to infection by the plant pathogen Pseudomonas syringae pv. tomato DC3000. Driven by the constitutive CaMV35S promoter, the cacao TcNPR1 gene partially complemented the npr1 mutation in transgenic Arabidopsis plants, resulting in 100 fold less bacterial growth in a leaf infection assay. Upon induction with SA, TcNPR1 was shown to translocate into the nucleus of leaf and root cells in a manner identical to Arabidopsis NPR1. Cacao NPR1 was also capable of participating in SA-JA signaling crosstalk, as evidenced by the suppression of JA responsive gene expression in TcNPR1 overexpressing transgenic plants. Conclusion Our data indicate that the TcNPR1 is a functional ortholog of Arabidopsis NPR1, and is likely to play a major role in defense response in cacao. This fundamental knowledge can contribute to breeding of disease resistant cacao varieties through the application of molecular markers or the use of transgenic strategies. PMID:21078185

Functional analysis of the Theobroma cacao NPR1 gene in Arabidopsis.

PubMed

Shi, Zi; Maximova, Siela N; Liu, Yi; Verica, Joseph; Guiltinan, Mark J

2010-11-15

The Arabidopsis thaliana NPR1 gene encodes a transcription coactivator (NPR1) that plays a major role in the mechanisms regulating plant defense response. After pathogen infection and in response to salicylic acid (SA) accumulation, NPR1 translocates from the cytoplasm into the nucleus where it interacts with other transcription factors resulting in increased expression of over 2000 plant defense genes contributing to a pathogen resistance response. A putative Theobroma cacao NPR1 cDNA was isolated by RT-PCR using degenerate primers based on homologous sequences from Brassica, Arabidopsis and Carica papaya. The cDNA was used to isolate a genomic clone from Theobroma cacao containing a putative TcNPR1 gene. DNA sequencing revealed the presence of a 4.5 kb coding region containing three introns and encoding a polypeptide of 591 amino acids. The predicted TcNPR1 protein shares 55% identity and 78% similarity to Arabidopsis NPR1, and contains each of the highly conserved functional domains indicative of this class of transcription factors (BTB/POZ and ankyrin repeat protein-protein interaction domains and a nuclear localization sequence (NLS)). To functionally define the TcNPR1 gene, we transferred TcNPR1 into an Arabidopsis npr1 mutant that is highly susceptible to infection by the plant pathogen Pseudomonas syringae pv. tomato DC3000. Driven by the constitutive CaMV35S promoter, the cacao TcNPR1 gene partially complemented the npr1 mutation in transgenic Arabidopsis plants, resulting in 100 fold less bacterial growth in a leaf infection assay. Upon induction with SA, TcNPR1 was shown to translocate into the nucleus of leaf and root cells in a manner identical to Arabidopsis NPR1. Cacao NPR1 was also capable of participating in SA-JA signaling crosstalk, as evidenced by the suppression of JA responsive gene expression in TcNPR1 overexpressing transgenic plants. Our data indicate that the TcNPR1 is a functional ortholog of Arabidopsis NPR1, and is likely to play a major role in defense response in cacao. This fundamental knowledge can contribute to breeding of disease resistant cacao varieties through the application of molecular markers or the use of transgenic strategies.

The drought response of Theobroma cacao (cacao) and the regulation of genes involved in polyamine biosynthesis by drought and other stresses.

PubMed

Bae, Hanhong; Kim, Soo-Hyung; Kim, Moon S; Sicher, Richard C; Lary, David; Strem, Mary D; Natarajan, Savithiry; Bailey, Bryan A

2008-02-01

Drought can negatively impact pod production despite the fact that cacao production usually occurs in tropical areas having high rainfall. Polyamines (PAs) have been associated with the response of plants to drought in addition to their roles in responses to many other stresses. The constitutive and drought inducible expression patterns of genes encoding enzymes involved in PA biosynthesis were determined: an ornithine decarboxylase (TcODC), an arginine decarboxylase (TcADC), an S-adenosylmethionine decarboxylase (TcSAMDC), a spermidine synthase (TcSPDS), and a spermine synthase (TcSPMS). Expression analysis using quantitative real-time reverse transcription-PCR (QPCR) results showed that the PA biosynthesis genes were expressed in all plant tissues examined. Constitutive expression of PA biosynthesis genes was generally highest in mature leaves and open flowers. Expression of TcODC, TcADC, and TcSAMDC was induced with the onset of drought and correlated with changes in stomatal conductance, photosynthesis, photosystem II efficiency, leaf water potential and altered emission of blue-green fluorescence from cacao leaves. Induction of TcSAMDC in leaves was most closely correlated with changes in water potential. The earliest measured responses to drought were enhanced expression of TcADC and TcSAMDC in roots along with decreases in stomatal conductance, photosynthesis, and photosystem II efficiency. Elevated levels of putrescine, spermidine, and spermine were detected in cacao leaves 13days after the onset of drought. Expression of all five PA associated transcripts was enhanced (1.5-3-fold) in response to treatment with abscisic acid. TcODC and TcADC, were also responsive to mechanical wounding, infection by Phytophthora megakarya (a causal agent of black pod disease in cacao), the necrosis- and ethylene-inducing protein (Nep1) of Fusarium oxysporum, and flower abscission. TcSAMDC expression was responsive to all stresses except flower abscission. TcODC, although constitutively expressed at much lower levels than TcADC, TcSAMDC, TcSPDS, and TcSPMS, was highly inducible by the fungal protein Nep1 (135-fold) and the cacao pathogen Phytophthora megakarya (671-fold). The full length cDNA for ODC was cloned and characterized. Among the genes studied, TcODC, TcADC, and TcSAMDC were most sensitive to induction by drought in addition to other abiotic and biotic stresses. TcODC, TcADC, and TcSAMDC may share signal transduction pathways and/or the stress induced signal induction pathways may converge at these three genes leading to similar although not identical patterns of expression. It is possible altering PA levels in cacao will result in enhanced tolerance to multiple stresses including drought and disease as has been demonstrated in other crops.

Isolation and identification of mycoparasitic isolates of Trichoderma asperellum with potential for suppression of black pod disease of cacao in Cameroon

USDA-ARS?s Scientific Manuscript database

Alternative measures to chemical fungicides are needed to control Phytophthora megakarya, the main causal agent of black pod diseasein Central and West Africa. Precolonized plate and detached cacao pod assays were used to screen fungal isolates for mycoparasitismon P. megakarya. Of over 200 isolates...

Cacao single-nucleotide polymorphism (SNP) markers: A discovery strategy to identify SNPs for genotyping, genetic mapping and genome wide association studies (GWAS)

USDA-ARS?s Scientific Manuscript database

Single-nucleotide polymorphisms (SNPs) are the most common genetic markers in Theobroma cacao, occurring approximately once in every 200 nucleotides. SNPs, like microsatellites, are co-dominant and PCR-based, but they have several advantages over microsatellites. They are unambiguous, so that a SN...

Atributos físicos do solo para a cultura do cacaueiro [Physical attributes of soil for the culture of cacao] (In Portuguese)

USDA-ARS?s Scientific Manuscript database

Cacao has achieved great importance due to its globally consumed products such as chocolate. In addition to the environmental benefits, since this culture is mainly managed in agroforestry systems, contributing to climate change mitigation. However, the assessment of soils physical attributes for ef...

Towards The Identification Of Candidate Genes Involved In Witches' Broom Disease Resistance In Theobroma cacao L.

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the source of cocoa beans for chocolate, is an important tropical agriculture commodity that is affected by a number of fungal pathogens and insect pests, as well as concerns about yield and quality. We are trying to find molecular genetic markers that are linked to disease resista...

Molecular and metabolic changes of cherelle wilt of cacao and its effect on Moniliophthora roreri

USDA-ARS?s Scientific Manuscript database

The seeds of Theobroma cacao L. pods are processed into cocoa products. Cherelle wilt is physiological thinning of young pods that result in loss of potential pods. Cherelle wilt first occurs 50 days after pollination (DAP) and a second thinning occurs around 70 DAP. Cherelles are also highly sus...

The Oil of Matico (Piper aduncum L.) an Alternative for the Control of Cacao Frosty Pod Rot (Moniliophthora roreri) in Peru

USDA-ARS?s Scientific Manuscript database

The cacao production in many Latin American countries is significantly reduced by frosty pod rot disease (Moniliophthora roreri) and yield reductions are to the extent of over 90% in many cases. The strategies of control includes: phytosanitation, genetic resistance, chemical and biological control....

Endophytic Trichoderma isolates from tropical environments delay disease onset and induce resistance against Phytophthora capsici in hot pepper using multiple mechanisms

USDA-ARS?s Scientific Manuscript database

Isolates of several Trichoderma spp., were collected from tropical environments as potential biocontrol agents for cacao (Theobroma cacao) diseases. The diversity of isolates collected, including new species, and there endophytic nature on their host plants, led us to consider if these isolates have...

Photosynthetic, antioxidative, molecular and ultrastructural responses of young cacao plants to Cd toxicity in the soil

USDA-ARS?s Scientific Manuscript database

Cadmium (Cd) is a highly toxic metal for plants, even at low concentrations in the soil. Humans and animals by ingesting plants, fruits, seeds or their derivatives contaminated with Cd can develop a number of chronic diseases, including cancer. The annual production of world cacao beans is approxima...

[Sexuality, heart and chocolate].

PubMed

Bianchi-Demicheli, F; Sekoranja, L; Pechère-Bertschi, A

2013-03-20

All along the history, many kinds of magic and aphrodisiac properties were attributed to the chocolate. Because of the presence of certain active substances, cacao and chocolate are supposed to have some potentially beneficial effects on human health, particularly on cardiovascular system. Containing flavoniods, cacao and its products have antioxidant, anti-inflammatory, anti-atherogenic, anti-thrombotic, antihypertensive and neuroprotective effects, as well as influence on insulin sensitivity, vascular endothelial function, and activation of nitric oxide. Other molecules, like methyxantin, biogenic amines and cannabinoid-like fatty acids, may have a psychoactive action. Synergic effect of all these substances could have a positive direct and indirect influence on sexual health and function. Nevertheless, randomized studies are needed to confirm these hypotheses and to elaborate recommendations about cacao consumption.

Efficient method of protein extraction from Theobroma cacao L. roots for two-dimensional gel electrophoresis and mass spectrometry analyses.

PubMed

Bertolde, F Z; Almeida, A-A F; Silva, F A C; Oliveira, T M; Pirovani, C P

2014-07-04

Theobroma cacao is a woody and recalcitrant plant with a very high level of interfering compounds. Standard protocols for protein extraction were proposed for various types of samples, but the presence of interfering compounds in many samples prevented the isolation of proteins suitable for two-dimensional gel electrophoresis (2-DE). An efficient method to extract root proteins for 2-DE was established to overcome these problems. The main features of this protocol are: i) precipitation with trichloroacetic acid/acetone overnight to prepare the acetone dry powder (ADP), ii) several additional steps of sonication in the ADP preparation and extractions with dense sodium dodecyl sulfate and phenol, and iii) adding two stages of phenol extractions. Proteins were extracted from roots using this new protocol (Method B) and a protocol described in the literature for T. cacao leaves and meristems (Method A). Using these methods, we obtained a protein yield of about 0.7 and 2.5 mg per 1.0 g lyophilized root, and a total of 60 and 400 spots could be separated, respectively. Through Method B, it was possible to isolate high-quality protein and a high yield of roots from T. cacao for high-quality 2-DE gels. To demonstrate the quality of the extracted proteins from roots of T. cacao using Method B, several protein spots were cut from the 2-DE gels, analyzed by tandem mass spectrometry, and identified. Method B was further tested on Citrus roots, with a protein yield of about 2.7 mg per 1.0 g lyophilized root and 800 detected spots.

Genes acquired by horizontal transfer are potentially involved in the evolution of phytopathogenicity in Moniliophthora perniciosa and Moniliophthora roreri, two of the major pathogens of cacao

USDA-ARS?s Scientific Manuscript database

Moniliophthora perniciosa and Moniliophthora roreri are phytopathogenic basidiomycete species that infect cacao and cause the two main diseases in this crop: “Witches’ Broom” and “Frosty Pod”, respectively. The ability of species from this genus (Moniliophthora) to cause disease is exceptional in th...

The differential influence of temperature on Phytophthora megakarya and Phytophthora palmivora pod lesion expansion, mycelia growth, gene expression, and metabolite profiles

USDA-ARS?s Scientific Manuscript database

Phytophthora megakarya and Phytophthora palmivora cause cacao black pod rot of cacao. P. megakarya occurs in Africa while P. palmivora is distributed world-wide. P. palmivora has a higher temperature maximum (34°C) than P. megakarya (30°C). Factors contributing to temperature maxima in Phytophtho...

Cacao biotechnology: current status and future prospects.

PubMed

Wickramasuriya, Anushka M; Dunwell, Jim M

2018-01-01

Theobroma cacao-The Food of the Gods, provides the raw material for the multibillion dollar chocolate industry and is also the main source of income for about 6 million smallholders around the world. Additionally, cocoa beans have a number of other nonfood uses in the pharmaceutical and cosmetic industries. Specifically, the potential health benefits of cocoa have received increasing attention as it is rich in polyphenols, particularly flavonoids. At present, the demand for cocoa and cocoa-based products in Asia is growing particularly rapidly and chocolate manufacturers are increasing investment in this region. However, in many Asian countries, cocoa production is hampered due to many reasons including technological, political and socio-economic issues. This review provides an overview of the present status of global cocoa production and recent advances in biotechnological applications for cacao improvement, with special emphasis on genetics/genomics, in vitro embryogenesis and genetic transformation. In addition, in order to obtain an insight into the latest innovations in the commercial sector, a survey was conducted on granted patents relating to T. cacao biotechnology. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Diversity of chloroplast genome among local clones of cocoa (Theobroma cacao, L.) from Central Sulawesi

NASA Astrophysics Data System (ADS)

Suwastika, I. Nengah; Pakawaru, Nurul Aisyah; Rifka, Rahmansyah, Muslimin, Ishizaki, Yoko; Cruz, André Freire; Basri, Zainuddin; Shiina, Takashi

2017-02-01

Chloroplast genomes typically range in size from 120 to 170 kilo base pairs (kb), which relatively conserved among plant species. Recent evaluation on several species, certain unique regions showed high variability which can be utilized in the phylogenetic analysis. Many fragments of coding regions, introns, and intergenic spacers, such as atpB-rbcL, ndhF, rbcL, rpl16, trnH-psbA, trnL-F, trnS-G, etc., have been used for phylogenetic reconstructions at various taxonomic levels. Based on that status, we would like to analysis the diversity of chloroplast genome within species of local cacao (Theobroma cacao L.) from Central Sulawesi. Our recent data showed, there were more than 20 clones from local farming in Central Sulawesi, and it can be detected based on phenotypic and nuclear-genome-based characterization (RAPD- Random Amplified Polymorphic DNA and SSR- Simple Sequences Repeat) markers. In developing DNA marker for this local cacao, here we also included analysis based on the variation of chloroplast genome. At least several regions such as rpl32-TurnL, it can be considered as chloroplast markers on our local clone of cocoa. Furthermore, we could develop phylogenetic analysis in between clones of cocoa.

The Genome of the Most Widely Cultivated Cacao Type and Its Use to Identify Candidate Genes Regulating Traits: Pod Color as an Example

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the source of cocoa beans for chocolate, is an important tropical agriculture commodity that is affected by a number of fungal pathogens and insect pests, as well as concerns about yield and quality. We are trying to find molecular genetic markers that are linked to disease resista...

Altered physiology, cell structure and gene expression of Theobroma cacao seedlings submitted to Cu toxicity

USDA-ARS?s Scientific Manuscript database

Theobroma cacao seedlings from the genotype CCN 51 were grown under greenhouse conditions and exposed to increasing concentrations of Cu (0.005, 1, 2, 4, 8, 16 and 32 mg Cu L-1) in nutrient solution. When doses were equal or higher than 8 mg Cu L-1, after 24 h of treatment application, leaf gas exch...

Diallel Analysis and Growth Parameters as Selection Tools for Drought Tolerance in Young Theobroma cacao Plants

PubMed Central

dos Santos, Emerson Alves; de Almeida, Alex-Alan Furtado; Ahnert, Dario; Branco, Marcia Christina da Silva; Valle, Raúl René; Baligar, Virupax C.

2016-01-01

This study aimed to estimate the combining ability, of T. cacao genotypes preselected for drought tolerance through diallel crosses. The experiment was conducted under greenhouse conditions at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a completely randomized block design, in an experimental arrangement 21 x 2 [21 complete diallel crosses and two water regimes (control and stressed)]. In the control, soil moisture was kept close to field capacity, with predawn leaf water potential (ΨWL) ranging from -0.1 to -0.5 MPa. In the drought regime, the soil moisture was reduced gradually by decreasing the amount of water application until ΨWL reached -2.0 to -2.5 MPa. Significant differences (p < 0.05) were observed for most morphological attributes analyzed regarding progenies, water regime and their interactions. The results of the joint diallel analysis revealed significant effects between general combining ability (GCA) x water regimes and between specific combining ability (SCA) x water regimes. The SCA 6 genetic material showed high general combining ability for growth variables regardless of the water regime. In general, the water deficit influenced the production of biomass in most of the evaluated T. cacao crosses, except for SCA-6 x IMC-67, Catongo x SCA, MOC-01 x Catongo, Catongo x IMC-67 and RB-40 x Catongo. Multivariate analysis showed that stem diameter (CD), total leaf area (TLA), leaf dry biomass (LDB), stem dry biomass (SDB), root dry biomass (RDB), total dry biomass (TDB), root length (RL), root volume (RV), root diameter (RD) <1 mm and 1 <(RD) <2 mm were the most important growth parameters in the separation of T. cacao genotypes in to tolerant and intolerant to soil water deficit. PMID:27504627

Diallel Analysis and Growth Parameters as Selection Tools for Drought Tolerance in Young Theobroma cacao Plants.

PubMed

Dos Santos, Emerson Alves; Almeida, Alex-Alan Furtado de; Ahnert, Dario; Branco, Marcia Christina da Silva; Valle, Raúl René; Baligar, Virupax C

2016-01-01

This study aimed to estimate the combining ability, of T. cacao genotypes preselected for drought tolerance through diallel crosses. The experiment was conducted under greenhouse conditions at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a completely randomized block design, in an experimental arrangement 21 x 2 [21 complete diallel crosses and two water regimes (control and stressed)]. In the control, soil moisture was kept close to field capacity, with predawn leaf water potential (ΨWL) ranging from -0.1 to -0.5 MPa. In the drought regime, the soil moisture was reduced gradually by decreasing the amount of water application until ΨWL reached -2.0 to -2.5 MPa. Significant differences (p < 0.05) were observed for most morphological attributes analyzed regarding progenies, water regime and their interactions. The results of the joint diallel analysis revealed significant effects between general combining ability (GCA) x water regimes and between specific combining ability (SCA) x water regimes. The SCA 6 genetic material showed high general combining ability for growth variables regardless of the water regime. In general, the water deficit influenced the production of biomass in most of the evaluated T. cacao crosses, except for SCA-6 x IMC-67, Catongo x SCA, MOC-01 x Catongo, Catongo x IMC-67 and RB-40 x Catongo. Multivariate analysis showed that stem diameter (CD), total leaf area (TLA), leaf dry biomass (LDB), stem dry biomass (SDB), root dry biomass (RDB), total dry biomass (TDB), root length (RL), root volume (RV), root diameter (RD) <1 mm and 1 <(RD) <2 mm were the most important growth parameters in the separation of T. cacao genotypes in to tolerant and intolerant to soil water deficit.

A genome survey of Moniliophthora perniciosa gives new insights into Witches' Broom Disease of cacao

PubMed Central

Mondego, Jorge MC; Carazzolle, Marcelo F; Costa, Gustavo GL; Formighieri, Eduardo F; Parizzi, Lucas P; Rincones, Johana; Cotomacci, Carolina; Carraro, Dirce M; Cunha, Anderson F; Carrer, Helaine; Vidal, Ramon O; Estrela, Raíssa C; García, Odalys; Thomazella, Daniela PT; de Oliveira, Bruno V; Pires, Acássia BL; Rio, Maria Carolina S; Araújo, Marcos Renato R; de Moraes, Marcos H; Castro, Luis AB; Gramacho, Karina P; Gonçalves, Marilda S; Neto, José P Moura; Neto, Aristóteles Góes; Barbosa, Luciana V; Guiltinan, Mark J; Bailey, Bryan A; Meinhardt, Lyndel W; Cascardo, Julio CM; Pereira, Gonçalo AG

2008-01-01

Background The basidiomycete fungus Moniliophthora perniciosa is the causal agent of Witches' Broom Disease (WBD) in cacao (Theobroma cacao). It is a hemibiotrophic pathogen that colonizes the apoplast of cacao's meristematic tissues as a biotrophic pathogen, switching to a saprotrophic lifestyle during later stages of infection. M. perniciosa, together with the related species M. roreri, are pathogens of aerial parts of the plant, an uncommon characteristic in the order Agaricales. A genome survey (1.9× coverage) of M. perniciosa was analyzed to evaluate the overall gene content of this phytopathogen. Results Genes encoding proteins involved in retrotransposition, reactive oxygen species (ROS) resistance, drug efflux transport and cell wall degradation were identified. The great number of genes encoding cytochrome P450 monooxygenases (1.15% of gene models) indicates that M. perniciosa has a great potential for detoxification, production of toxins and hormones; which may confer a high adaptive ability to the fungus. We have also discovered new genes encoding putative secreted polypeptides rich in cysteine, as well as genes related to methylotrophy and plant hormone biosynthesis (gibberellin and auxin). Analysis of gene families indicated that M. perniciosa have similar amounts of carboxylesterases and repertoires of plant cell wall degrading enzymes as other hemibiotrophic fungi. In addition, an approach for normalization of gene family data using incomplete genome data was developed and applied in M. perniciosa genome survey. Conclusion This genome survey gives an overview of the M. perniciosa genome, and reveals that a significant portion is involved in stress adaptation and plant necrosis, two necessary characteristics for a hemibiotrophic fungus to fulfill its infection cycle. Our analysis provides new evidence revealing potential adaptive traits that may play major roles in the mechanisms of pathogenicity in the M. perniciosa/cacao pathosystem. PMID:19019209

Theobroma cacao increases cells viability and reduces IL-6 and sVCAM-1 level in endothelial cells induced by plasma from preeclamptic patients.

PubMed

Rahayu, Budi; Baktiyani, Siti Candra Windu; Nurdiana, Nurdiana

2016-01-01

This study aims to investigate whether an ethanolic extract of Theobroma cacao bean is able to increase cell viability and decrease IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Endothelial cells were obtained from human umbilical vascular endothelial cells. At confluency, endothelial cells were divided into six groups, which included control (untreated), endothelial cells exposed to plasma from normal pregnancy, endothelial cells exposed to 2% plasma from preeclamptic patients (PP), endothelial cells exposed to PP in the presence of ethanolic extract of T. cacao (PP+TC) at the following three doses: 25, 50, and 100 ppm. The analysis was performed in silico using the Hex 8.0, LigPlus and LigandScout 3.1 software. Analysis on IL-6 and sVCAM-1 levels were done by enzyme linked immunosorbent assay (ELISA). We found that seven of them could bind to the protein NFκB (catechin, leucoanthocyanidin, niacin, phenylethylamine, theobromine, theophylline, and thiamin). This increase in IL-6 was significantly (P<0.05) attenuated by both the 50 and 100 ppm treatments of T. cacao extract. Plasma from PP significantly increased sVCAM-1 levels compared to untreated cells. This increase in sVCAM-1 was significantly attenuated by all doses of the extract. In conclusion, T. cacao extract prohibits the increase in IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Therefore this may provide a herbal therapy for attenuating the endothelial dysfunction found in preeclampsia. Copyright © 2016 International Society for the Study of Hypertension in Pregnancy. Published by Elsevier B.V. All rights reserved.

Altered physiology, cell structure, and gene expression of Theobroma cacao seedlings subjected to Cu toxicity.

PubMed

Souza, Vânia L; de Almeida, Alex-Alan F; Souza, Jadiel de S; Mangabeira, Pedro A O; de Jesus, Raildo M; Pirovani, Carlos P; Ahnert, Dário; Baligar, Virupax C; Loguercio, Leandro L

2014-01-01

Seedlings of Theobroma cacao CCN 51 genotype were grown under greenhouse conditions and exposed to increasing concentrations of Cu (0.005, 1, 2, 4, 8, 16, and 32 mg Cu L(-1)) in nutrient solution. When doses were equal or higher than 8 mg Cu L(-1), after 24 h of treatment application, leaf gas exchange was highly affected and changes in chloroplasts thylakoids of leaf mesophyll cells and plasmolysis of cells from the root cortical region were observed. In addition, cell membranes of roots and leaves were damaged. In leaves, 96 h after treatments started, increases in the percentage of electrolyte leakage through membranes were observed with increases of Cu in the nutrient solution. Moreover, there was an increase in the concentration of thiobarbituric acid-reactive substances in roots due to lipid peroxidation of membranes. Chemical analysis showed that increases in Cu concentrations in vegetative organs of T. cacao increased with the increase of the metal in the nutrient solution, but there was a greater accumulation of Cu in roots than in shoots. The excess of Cu interfered in the levels of Mn, Zn, Fe, Mg, K, and Ca in different organs of T. cacao. Analysis of gene expression via RTq-PCR showed increased levels of MT2b, SODCyt, and PER-1 expression in roots and of MT2b, PSBA, PSBO, SODCyt, and SODChI in leaves. Hence, it was concluded that Cu in nutrient solution at doses equal or above 8 mg L(-1) significantly affected leaf gas exchange, cell ultrastructure, and transport of mineral nutrients in seedlings of this T. cacao genotype.

Differential gene expression by Moniliophthora roreri while overcoming cacao tolerance in the field.

PubMed

Bailey, Bryan A; Melnick, Rachel L; Strem, Mary D; Crozier, Jayne; Shao, Jonathan; Sicher, Richard; Phillips-Mora, Wilberth; Ali, Shahin S; Zhang, Dapeng; Meinhardt, Lyndel

2014-09-01

Frosty pod rot (FPR) of Theobroma cacao (cacao) is caused by the hemibiotrophic fungus Moniliophthora roreri. Cacao clones tolerant to FPR are being planted throughout Central America. To determine whether M. roreri shows a differential molecular response during successful infections of tolerant clones, we collected field-infected pods at all stages of symptomatology for two highly susceptible clones (Pound-7 and CATIE-1000) and three tolerant clones (UF-273, CATIE-R7 and CATIE-R4). Metabolite analysis was carried out on clones Pound-7, CATIE-1000, CATIE-R7 and CATIE-R4. As FPR progressed, the concentrations of sugars in pods dropped, whereas the levels of trehalose and mannitol increased. Associations between symptoms and fungal loads and some organic and amino acid concentrations varied depending on the clone. RNA-Seq analysis identified 873 M. roreri genes that were differentially expressed between clones, with the primary difference being whether the clone was susceptible or tolerant. Genes encoding transcription factors, heat shock proteins, transporters, enzymes modifying membranes or cell walls and metabolic enzymes, such as malate synthase and alternative oxidase, were differentially expressed. The differential expression between clones of 43 M. roreri genes was validated by real-time quantitative reverse transcription polymerase chain reaction. The expression profiles of some genes were similar in susceptible and tolerant clones (other than CATIE-R4) and varied with the biotrophic/necrotropic shift. Moniliophthora roreri genes associated with stress metabolism and responses to heat shock and anoxia were induced early in tolerant clones, their expression profiles resembling that of the necrotrophic phase. Moniliophthora roreri stress response genes, induced during the infection of tolerant clones, may benefit the fungus in overcoming cacao defense mechanisms. © 2014 BSPP AND JOHN WILEY & SONS LTD.

Genetic Parameters and the Impact of Off-Types for Theobroma cacao L. in a Breeding Program in Brazil

PubMed Central

DuVal, Ashley; Gezan, Salvador A.; Mustiga, Guiliana; Stack, Conrad; Marelli, Jean-Philippe; Chaparro, José; Livingstone, Donald; Royaert, Stefan; Motamayor, Juan C.

2017-01-01

Breeding programs of cacao (Theobroma cacao L.) trees share the many challenges of breeding long-living perennial crops, and genetic progress is further constrained by both the limited understanding of the inheritance of complex traits and the prevalence of technical issues, such as mislabeled individuals (off-types). To better understand the genetic architecture of cacao, in this study, 13 years of phenotypic data collected from four progeny trials in Bahia, Brazil were analyzed jointly in a multisite analysis. Three separate analyses (multisite, single site with and without off-types) were performed to estimate genetic parameters from statistical models fitted on nine important agronomic traits (yield, seed index, pod index, % healthy pods, % pods infected with witches broom, % of pods other loss, vegetative brooms, diameter, and tree height). Genetic parameters were estimated along with variance components and heritabilities from the multisite analysis, and a trial was fingerprinted with low-density SNP markers to determine the impact of off-types on estimations. Heritabilities ranged from 0.37 to 0.64 for yield and its components and from 0.03 to 0.16 for disease resistance traits. A weighted index was used to make selections for clonal evaluation, and breeding values estimated for the parental selection and estimation of genetic gain. The impact of off-types to breeding progress in cacao was assessed for the first time. Even when present at <5% of the total population, off-types altered selections by 48%, and impacted heritability estimations for all nine of the traits analyzed, including a 41% difference in estimated heritability for yield. These results show that in a mixed model analysis, even a low level of pedigree error can significantly alter estimations of genetic parameters and selections in a breeding program. PMID:29250097

Expression of the Theobroma cacao Bax-inhibitor-1 gene in tomato reduces infection by the hemibiotrophic pathogen Moniliophthora perniciosa.

PubMed

Scotton, Danielle Camargo; Azevedo, Mariana Da Silva; Sestari, Ivan; Da Silva, Jamille Santos; Souza, Lucas Anjos; Peres, Lázaro Eustáquio Pereira; Leal, Gildemberg Amorim; Figueira, Antonio

2017-10-01

Programmed cell death (PCD) plays a key role in plant responses to pathogens, determining the success of infection depending on the pathogen lifestyle and on which participant of the interaction triggers cell death. The hemibiotrophic basidiomycete Moniliophthora perniciosa is the causal agent of witches' broom disease of Theobroma cacao L. (cacao), a serious constraint for production in South America and the Caribbean. It has been hypothesized that M. perniciosa pathogenesis involves PCD, initially as a plant defence mechanism, which is diverted by the fungus to induce necrosis during the dikaryotic phase of the mycelia. Here, we evaluated whether the expression of a cacao anti-apoptotic gene would affect the incidence and severity of M. perniciosa infection using the 'Micro-Tom' (MT) tomato as a model. The cacao Bax-inhibitor-1 (TcBI-1) gene, encoding a putative basal attenuator of PCD, was constitutively expressed in MT to evaluate function. Transformants expressing TcBI-1, when treated with tunicamycin, an inducer of endoplasmic reticulum stress, showed a decrease in cell peroxidation. When the same transformants were inoculated with the necrotrophic fungal pathogens Sclerotinia sclerotiorum, Sclerotium rolfsii and Botrytis cinerea, a significant reduction in infection severity was observed, confirming TcBI-1 function. After inoculation with M. perniciosa, TcBI-1 transformant lines showed a significant reduction in disease incidence compared with MT. The overexpression of TcBI-1 appears to affect the ability of germinating spores to penetrate susceptible tissues, restoring part of the non-host resistance in MT against the S-biotype of M. perniciosa. © 2016 BSPP AND JOHN WILEY & SONS LTD.

Inducible somatic embryogenesis in Theobroma cacao achieved using the DEX-activatable transcription factor-glucocorticoid receptor fusion.

PubMed

Shires, Morgan E; Florez, Sergio L; Lai, Tina S; Curtis, Wayne R

2017-11-01

To carry out mass propagation of superior plants to improve agricultural and silvicultural production though advancements in plant cell totipotency, or the ability of differentiated somatic plant cells to regenerate an entire plant. The first demonstration of a titratable control over somatic embryo formation in a commercially relevant plant, Theobroma cacao (Chocolate tree), was achieved using a dexamethasone activatable chimeric transcription factor. This four-fold enhancement in embryo production rate utilized a glucocorticoid receptor fused to an embryogenic transcription factor LEAFY COTYLEDON 2. Where previous T. cacao somatic embryogenesis has been restricted to dissected flower parts, this construct confers an unprecedented embryogenic potential to leaves. Activatable chimeric transcription factors provide a means for elucidating the regulatory cascade associated with plant somatic embryogenesis towards improving its use for somatic regeneration of transgenics and plant propagation.

Towards the understanding of the cocoa transcriptome: Production and analysis of an exhaustive dataset of ESTs of Theobroma cacao L. generated from various tissues and under various conditions.

USDA-ARS?s Scientific Manuscript database

Theobroma cacao L., is a tree originated from the tropical rainforest of South America. It is one of the major cash crops for many tropical countries. Cocoa is mainly produced on small holdings, providing resources for 14 million farmers. Disease resistance and cocoa quality improvement are two impo...

Use of Bennett's Hierarchical Model in the Evaluation of the Extension Education Program for Cacao Farmers in the Northeast Region of the Dominican Republic. Summary of Research 54.

ERIC Educational Resources Information Center

De los Santos, Saturnino; Norland, Emmalou Van Tilburg

A study evaluated the cacao farmer training program in the Dominican Republic by testing hypothesized relationships among reactions, knowledge and skills, attitudes, aspirations, and some selected demographic characteristics of farmers who attended programs. Bennett's hierarchical model of program evaluation was used as the framework of the study.…

Cacao pod husks as a source of low-methoxyl, highly acetylated pectins able to gel in acidic media.

PubMed

Vriesmann, Lúcia Cristina; de Oliveira Petkowicz, Carmen Lúcia

2017-08-01

Cacao pod husks, the main by-product from cocoa production, have been investigated for pectin isolation. In the present study, the rheological properties of two low-methoxyl (LM) pectins isolated from cacao pod husks using different extraction conditions were evaluated. One pectin was obtained from optimized conditions employing aqueous nitric acid as an extractant, and the other one was extracted with boiling water. Pectin gels (0.99% galacturonic acid equivalent, w/w) were prepared at pH 2.5-3.0 in the presence of 60% sucrose (w/w) and subjected to rheological analysis. Dynamic oscillatory experiments at 25°C indicated that better gels were obtained at the lowest pH (2.5). Steady shear measurements revealed a shear-thinning behavior. The apparent viscosities of the samples increased as pH decreased. Gelation with calcium ions was not observed for either of the highly acetylated LM pectins analyzed. The rheological analysis results showed that despite their high acetyl content, LM pectins extracted by different methods from cacao pod husks were able to form gels at low pH under reduced water activity, suggesting a possible application in acidic products. Copyright © 2017 Elsevier B.V. All rights reserved.

High-affinity copper transport and Snq2 export permease of saccharomyces cerevisiae modulate cytotoxicity of PR-10 from Theobroma cacao.

PubMed

Pungartnik, Cristina; da Silva, Aline Clara; de Melo, Sarah Alves; Gramacho, Karina Peres; de Mattos Cascardo, Júlio Cézar; Brendel, Martin; Micheli, Fabienne; da Silva Gesteira, Abelmon

2009-01-01

A pathogenesis-related (PR) protein from Theobroma cacao (TcPR-10) was identified from a cacao-Moniliophthora perniciosa interaction cDNA library. Nucleotide and amino acid sequences showed homology with other PR-10 proteins having P loop motif and Betv1 domain. Recombinant TcPR-10 showed in vitro and in vivo ribonuclease activity, and antifungal activity against the basidiomycete cacao pathogen M. perniciosa and the yeast Saccharomyces cerevisiae. Fluorescein isothiocyanate-labeled TcPR-10 was internalized by M. perniciosa hyphae and S. cerevisiae cells and inhibited growth of both fungi. Energy and temperature-dependent internalization of the TcPR-10 suggested an active importation into the fungal cells. Chronical exposure to TcPR-10 of 29 yeast mutants with single gene defects in DNA repair, general membrane transport, metal transport, and antioxidant defenses was tested. Two yeast mutants were hyperresistant compared with their respective isogenic wild type: ctr3Delta mutant, lacking the high-affinity plasma membrane copper transporter and mac1Delta, the copper-sensing transcription factor involved in regulation of high-affinity copper transport. Acute exposure of exponentially growing yeast cells revealed that TcPR-10 resistance is also enhanced in the Snq2 export permease-lacking mutant which has reduced intracellular presence of TcPR-10.

Factors Related to Adoption and Non-Adoption of Technical and Organizational Recommendations by Farmers Involved with Societe de Developpement du Cacao (SO.DE.CAO) in Cameroon. A Research Summary.

ERIC Educational Resources Information Center

Kamga, Andre; Cheek, Jimmy G.

In order to promote cocoa production and assist cocoa farmers in overcoming diseases in this crop, the government of Cameroon created an experimental corporation called Societe de Developpement du Cacao (SO.DE.CAO) in 1974. This organization functioned much like an extension service to provide information about crop production and disease control.…

Theobroma cacao: Review of the Extraction, Isolation, and Bioassay of Its Potential Anti-cancer Compounds

PubMed Central

Baharum, Zainal; Akim, Abdah Md; Hin, Taufiq Yap Yun; Hamid, Roslida Abdul; Kasran, Rosmin

2016-01-01

Plants have been a good source of therapeutic agents for thousands of years; an impressive number of modern drugs used for treating human diseases are derived from natural sources. The Theobroma cacao tree, or cocoa, has recently garnered increasing attention and become the subject of research due to its antioxidant properties, which are related to potential anti-cancer effects. In the past few years, identifying and developing active compounds or extracts from the cocoa bean that might exert anti-cancer effects have become an important area of health- and biomedicine-related research. This review provides an updated overview of T. cacao in terms of its potential anti-cancer compounds and their extraction, in vitro bioassay, purification, and identification. This article also discusses the advantages and disadvantages of the techniques described and reviews the processes for future perspectives of analytical methods from the viewpoint of anti-cancer compound discovery. PMID:27019680

Analysis of community income on suren (Toona sureni (Blume) Merr.) and cacao crops (Theobroma cacao L.) in Simalungun, North Sumatera-Indonesia

NASA Astrophysics Data System (ADS)

Latifah, S.; Afifuddin, Y.; Widya, S.

2018-02-01

Agroforestry is the management and integration of trees, crops and/or livestock on the same plot of land and can be an integral component of productive agriculture. It may include existing native forests and forests established by landholders. The study was conducted in Mekar Sari Raya village, Panei sub, Simalungun regency, North Sumatera. This study aims to gain the ability to use agroforestry in suren crops and cocoa that provides benefits to farmers and the feasibility of the model farm. The study site has Net Present Value (NPV) is 2,670,306,905 ( IDR) for 15 of year, Gross B/C Ratio (BCR) is 2.3; Internal Rate of Return ( IRR) is 28 %; and Payback Period (PP) for 5 years 4 months 24 days. Agroforestry using commodities cacao and suren crops are financially feasible to be cultivated and developed.

Crystal Structure of MpPR-1i, a SCP/TAPS protein from Moniliophthora perniciosa, the fungus that causes Witches’ Broom Disease of Cacao

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baroni, Renata M.; Luo, Zhipu; Darwiche, Rabih

The pathogenic fungi Moniliophthora perniciosa causes Witches’ Broom Disease (WBD) of cacao. The structure of MpPR-1i, a protein expressed by M. perniciosa when it infects cacao, are presented. This is the first reported de novo structure determined by single-wavelength anomalous dispersion phasing upon soaking with selenourea. Each monomer has flexible loop regions linking the core alpha-beta-alpha sandwich topology that comprise ~50% of the structure, making it difficult to generate an accurate homology model of the protein. MpPR-1i is monomeric in solution but is packed as a high ~70% solvent content, crystallographic heptamer. The greatest conformational flexibility between monomers is foundmore » in loops exposed to the solvent channel that connect the two longest strands. MpPR-1i lacks the conserved CAP tetrad and is incapable of binding divalent cations. MpPR-1i has the ability to bind lipids, which may have roles in its infection of cacao. These lipids likely bind in the palmitate binding cavity as observed in tablysin-15, since MpPR-1i binds palmitate with comparable affinity as tablysin-15. Further studies are required to clarify the possible roles and underlying mechanisms of neutral lipid binding, as well as their effects on the pathogenesis of M. perniciosa so as to develop new interventions for WBD.« less

In vitro antioxidant and antiproliferative activities of methanolic plant part extracts of Theobroma cacao.

PubMed

Baharum, Zainal; Akim, Abdah Md; Taufiq-Yap, Yun Hin; Hamid, Roslida Abdul; Kasran, Rosmin

2014-11-10

The aims of this study were to determine the antioxidant and antiproliferative activity of the following Theobroma cacao plant part methanolic extracts: leaf, bark, husk, fermented and unfermented shell, pith, root, and cherelle. Antioxidant activity was determined using 2,2-diphenyl-2-picrylhydrazyl (DPPH), thiobarbituric acid-reactive substances (TBARS), and Folin-Ciocalteu assays; the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium (MTT) assay was used to determine antiproliferative activity. The root extract had the highest antioxidant activity; its median effective dose (EC50) was 358.3±7.0 µg/mL and total phenolic content was 22.0±1.1 g GAE/100 g extract as compared to the other methanolic plant part extracts. Only the cherelle extract demonstrated 10.4%±1.1% inhibition activity in the lipid peroxidation assay. The MTT assay revealed that the leaf extract had the highest antiproliferative activity against MCF-7 cells [median inhibitory concentration (IC50)=41.4±3.3 µg/mL]. Given the overall high IC50 for the normal liver cell line WRL-68, this study indicates that T. cacao methanolic extracts have a cytotoxic effect in cancer cells, but not in normal cells. Planned future investigations will involve the purification, identification, determination of the mechanisms of action, and molecular assay of T. cacao plant extracts.

Apoplastic and intracellular plant sugars regulate developmental transitions in witches’ broom disease of cacao

PubMed Central

Barau, Joan; Grandis, Adriana; Carvalho, Vinicius Miessler de Andrade; Teixeira, Gleidson Silva; Zaparoli, Gustavo Henrique Alcalá; do Rio, Maria Carolina Scatolin; Rincones, Johana; Buckeridge, Marcos Silveira; Pereira, Gonçalo Amarante Guimarães

2015-01-01

Witches’ broom disease (WBD) of cacao differs from other typical hemibiotrophic plant diseases by its unusually long biotrophic phase. Plant carbon sources have been proposed to regulate WBD developmental transitions; however, nothing is known about their availability at the plant–fungus interface, the apoplastic fluid of cacao. Data are provided supporting a role for the dynamics of soluble carbon in the apoplastic fluid in prompting the end of the biotrophic phase of infection. Carbon depletion and the consequent fungal sensing of starvation were identified as key signalling factors at the apoplast. MpNEP2, a fungal effector of host necrosis, was found to be up-regulated in an autophagic-like response to carbon starvation in vitro. In addition, the in vivo artificial manipulation of carbon availability in the apoplastic fluid considerably modulated both its expression and plant necrosis rate. Strikingly, infected cacao tissues accumulated intracellular hexoses, and showed stunted photosynthesis and the up-regulation of senescence markers immediately prior to the transition to the necrotrophic phase. These opposite findings of carbon depletion and accumulation in different host cell compartments are discussed within the frame of WBD development. A model is suggested to explain phase transition as a synergic outcome of fungal-related factors released upon sensing of extracellular carbon starvation, and an early senescence of infected tissues probably triggered by intracellular sugar accumulation. PMID:25540440

Establishing references for gene expression analyses by RT-qPCR in Theobroma cacao tissues.

PubMed

Pinheiro, T T; Litholdo, C G; Sereno, M L; Leal, G A; Albuquerque, P S B; Figueira, A

2011-11-17

Lack of continuous progress in Theobroma cacao (Malvaceae) breeding, especially associated with seed quality traits, requires more efficient selection methods based on genomic information. Reverse transcript quantitative PCR (RT-qPCR) has become the method of choice for gene expression analysis, but relative expression analysis requires various reference genes, which must be stable across various biological conditions. We sought suitable reference genes for various tissues of cacao, especially developing seeds. Ten potential reference genes were analyzed for stability at various stages of embryo development, leaves, stems, roots, flowers, and pod epicarp; seven of them were also evaluated in shoot tips treated either with hormones (salicylate; ethefon; methyl-jasmonate) or after inoculation with the fungus Moniliophthora perniciosa (Marasmiaceae sensu lato). For developing embryos, the three most stable genes were actin (ACT), polyubiquitin (PUB), and ribosomal protein L35 (Rpl35). In the analyses of various tissues, the most stable genes were malate dehydrogenase (MDH), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and acyl-carrier protein B (ACP B). GAPDH, MDH and tubulin (TUB) were the most appropriate for normalization when shoot apexes were treated with hormones, while ACT, TUB and Rpl35 were the most appropriate after inoculation with M. perniciosa. We conclude that for each plant system and biological or ontogenetical condition, there is a need to define suitable reference genes. This is the first report to define reference genes for expression studies in cacao.

Genetic Population Structure of Cacao Plantings within a Young Production Area in Nicaragua

PubMed Central

Trognitz, Bodo; Scheldeman, Xavier; Hansel-Hohl, Karin; Kuant, Aldo; Grebe, Hans; Hermann, Michael

2011-01-01

Significant cocoa production in the municipality of Waslala, Nicaragua, began in 1961. Since the 1980s, its economic importance to rural smallholders increased, and the region now contributes more than 50% of national cocoa bean production. This research aimed to assist local farmers to develop production of high-value cocoa based on optimal use of cacao biodiversity. Using microsatellite markers, the allelic composition and genetic structure of cacao was assessed from 44 representative plantings and two unmanaged trees. The population at Waslala consists of only three putative founder genotype spectra (lineages). Two (B and R) were introduced during the past 50 years and occur in >95% of all trees sampled, indicating high rates of outcrossing. Based on intermediate allelic diversity, there was large farm-to-farm multilocus genotypic variation. GIS analysis revealed unequal distribution of the genotype spectra, with R being frequent within a 2 km corridor along roads, and B at more remote sites with lower precipitation. The third lineage, Y, was detected in the two forest trees. For explaining the spatial stratification of the genotype spectra, both human intervention and a combination of management and selection driven by environmental conditions, appear responsible. Genotypes of individual trees were highly diverse across plantings, thus enabling selection for farm-specific qualities. On-farm populations can currently be most clearly recognized by the degree of the contribution of the three genotype spectra. Of two possible strategies for future development of cacao in Waslala, i.e. introducing more unrelated germplasm, or working with existing on-site diversity, the latter seems most appropriate. Superior genotypes could be selected by their specific composite genotype spectra as soon as associations with desired quality traits are established, and clonally multiplied. The two Y trees from the forest share a single multilocus genotype, possibly representing the Mayan, ‘ancient Criollo’ cacao. PMID:21264251

Drought effects on soil CO

NASA Astrophysics Data System (ADS)

van Straaten, O.; Veldkamp, E.; Köhler, M.; Anas, I.

2009-12-01

Climate change induced droughts pose a serious threat to ecosystems across the tropics and sub-tropics, particularly to those areas not adapted to natural dry periods. In order to study the vulnerability of cacao (Theobroma cacao) - Gliricidia sepium agroforestry plantations to droughts a large scale throughfall displacement roof was built in Central Sulawesi, Indonesia. In this 19-month replicated experiment, we measured soil surface CO2 efflux (soil respiration) in three simulated drought plots compared with three adjacent control plots. Soil respiration rates peaked at intermediate soil moisture and decreased under increasingly dry conditions (drought induced), but also decreased when soils became water saturated, as evidenced in control plots. The simulated drought plots exhibited a slight decrease in soil respiration compared to the control plots (average 13% decrease). The strength of the drought effect was spatially variable - while some measurement chamber sites reacted strongly ("responsive") to the decrease in soil water content (up to R2=0.70) (n=11), others did not react at all ("non-responsive") (n=7). The degree of soil CO2 respiration drought response was highest around cacao tree stems and decreased with distance from the stem (R2=0.22). A significant correlation was measured between "responsive" soil respiration chamber sites and sap flux density ratios of cacao (R=0.61) and Gliricidia (R=0.65). Leaf litter CO2 respiration decreased as conditions became drier. During dry periods the litter layer contributed approximately 3-4% of the total CO2 efflux and up to 40% during wet periods. A CO2 flush was recorded during the rewetting phase that lasted for approximately two weeks, during which time accumulated labile carbon stocks mineralized. The net effect on soil CO2 emissions over the duration of the experiment was neutral, control plots respired 11.1±0.5 Mg C ha-1 yr-1, while roof plots respired 10.5±0.5 Mg C ha-1 yr-1.

Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease.

PubMed

Camillo, Luciana Rodrigues; Filadelfo, Ciro Ribeiro; Monzani, Paulo Sérgio; Corrêa, Ronan Xavier; Gramacho, Karina Peres; Micheli, Fabienne; Pirovani, Carlos Priminho

2013-12-01

The level of hydrogen peroxide (H2O2) in plants signalizes the induction of several genes, including that of ascorbate peroxidase (APX-EC 1.11.1.11). APX isoenzymes play a central role in the elimination of intracellular H2O2 and contribute to plant responses to diverse stresses. During the infection process in Theobroma cacao by Moniliophthora perniciosa oxidative stress is generated and the APX action recruited from the plant. The present work aimed to characterize the T. cacao APX involved in the molecular interaction of T. cacao-M. perniciosa. The peroxidase activity was analyzed in protein extracts from cocoa plants infected by M. perniciosa and showed the induction of peroxidases like APX in resistant cocoa plants. The cytosolic protein of T. cacao (GenBank: ABR68691.2) was phylogenetically analyzed in relation to other peroxidases from the cocoa genome and eight genes encoding APX proteins with conserved domains were also analyzed. The cDNA from cytosolic APX was cloned in pET28a and the recombinant protein expressed and purified (rTc-cAPX). The secondary structure of the protein was analyzed by Circular Dichroism (CD) displaying high proportion of α-helices when folded. The enzymatic assay shows stable activity using ascorbate and guaiacol as an electron donor for H2O2 reduction. The pH 7.5 is the optimum for enzyme activity. Chromatographic analysis suggests that rTc-cAPX is a homodimer in solution. Results indicate that the rTc-cAPX is correctly folded, stable and biochemically active. The purified rTc-cAPX presented biotechnological potential and is adequate for future structural and functional studies. Copyright © 2013 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Genetic population structure of cacao plantings within a young production area in Nicaragua.

PubMed

Trognitz, Bodo; Scheldeman, Xavier; Hansel-Hohl, Karin; Kuant, Aldo; Grebe, Hans; Hermann, Michael

2011-01-14

Significant cocoa production in the municipality of Waslala, Nicaragua, began in 1961. Since the 1980s, its economic importance to rural smallholders increased, and the region now contributes more than 50% of national cocoa bean production. This research aimed to assist local farmers to develop production of high-value cocoa based on optimal use of cacao biodiversity. Using microsatellite markers, the allelic composition and genetic structure of cacao was assessed from 44 representative plantings and two unmanaged trees. The population at Waslala consists of only three putative founder genotype spectra (lineages). Two (B and R) were introduced during the past 50 years and occur in >95% of all trees sampled, indicating high rates of outcrossing. Based on intermediate allelic diversity, there was large farm-to-farm multilocus genotypic variation. GIS analysis revealed unequal distribution of the genotype spectra, with R being frequent within a 2 km corridor along roads, and B at more remote sites with lower precipitation. The third lineage, Y, was detected in the two forest trees. For explaining the spatial stratification of the genotype spectra, both human intervention and a combination of management and selection driven by environmental conditions, appear responsible. Genotypes of individual trees were highly diverse across plantings, thus enabling selection for farm-specific qualities. On-farm populations can currently be most clearly recognized by the degree of the contribution of the three genotype spectra. Of two possible strategies for future development of cacao in Waslala, i.e. introducing more unrelated germplasm, or working with existing on-site diversity, the latter seems most appropriate. Superior genotypes could be selected by their specific composite genotype spectra as soon as associations with desired quality traits are established, and clonally multiplied. The two Y trees from the forest share a single multilocus genotype, possibly representing the Mayan, 'ancient Criollo' cacao.

Endophytic Association of Trichoderma asperellum within Theobroma cacao Suppresses Vascular Streak Dieback Incidence and Promotes Side Graft Growth

PubMed Central

Nasaruddin, Nasaruddin; Hendarto, Hendarto; Hakkar, Andi Akbar; Agriansyah, Nursalim

2016-01-01

Trichoderma species are able to persist on living sapwood and leaves of cacao (Theobroma cacao) in an endophytic relationship. In this research, we evaluated the ability of Trichodema asperellum introduced at the incision site in the bark for side grafting with the concentration of 4 g/10 mL, 4 g/100 mL, and 4 g/1,000 mL (suspended in water) in suppressing vascular streak dieback (VSD) incidence and promoting growth of side grafts in the field. The incidence of VSD in two local clones of cacao, MCC1 and M04, without application of T. asperellum was 71.2% and 70.1% at 21 wk after grafting, respectively. However, when the two clones were treated with a concentration of 4 g/10 mL T. asperellum, the incidence was 20.6% and 21.7%, respectively, compared to 29.1% and 20.9% at 4 g/100 mL and 18.2% and 15.6% at 4 g/1,000 mL. By comparing to the control, the treatment with the same concentrations of T. asperellum listed above, the total number of stomata in MCC1 decreased by 41.9%, 30.2%, and 14.0% and in M04 by 30.5%, 21.9%, and -2.5% (exception), respectively. Otherwise, the total area of stomata opening increased by 91.4%, 99.7%, and 28.6% in MCC1 and by 203.8%, 253.5%, and 35.9% in M04, respectively. Furthermore, the number of buds and branches treated with a mixture concentration on the the two clones increased by 90.7% and 21.7%, respectively. These data showed that the application of T. asperellum to cacao scions while grafting can decrease VSD incidence in side grafts and increase growth of grafts in addition to decreasing total number of stomata, increasing total area of opened stomata, and increasing number of buds and branches. PMID:27790069

Endophytic Association of Trichoderma asperellum within Theobroma cacao Suppresses Vascular Streak Dieback Incidence and Promotes Side Graft Growth.

PubMed

Rosmana, Ade; Nasaruddin, Nasaruddin; Hendarto, Hendarto; Hakkar, Andi Akbar; Agriansyah, Nursalim

2016-09-01

Trichoderma species are able to persist on living sapwood and leaves of cacao ( Theobroma cacao ) in an endophytic relationship. In this research, we evaluated the ability of Trichodema asperellum introduced at the incision site in the bark for side grafting with the concentration of 4 g/10 mL, 4 g/100 mL, and 4 g/1,000 mL (suspended in water) in suppressing vascular streak dieback (VSD) incidence and promoting growth of side grafts in the field. The incidence of VSD in two local clones of cacao, MCC1 and M04, without application of T. asperellum was 71.2% and 70.1% at 21 wk after grafting, respectively. However, when the two clones were treated with a concentration of 4 g/10 mL T. asperellum , the incidence was 20.6% and 21.7%, respectively, compared to 29.1% and 20.9% at 4 g/100 mL and 18.2% and 15.6% at 4 g/1,000 mL. By comparing to the control, the treatment with the same concentrations of T. asperellum listed above, the total number of stomata in MCC1 decreased by 41.9%, 30.2%, and 14.0% and in M04 by 30.5%, 21.9%, and -2.5% (exception), respectively. Otherwise, the total area of stomata opening increased by 91.4%, 99.7%, and 28.6% in MCC1 and by 203.8%, 253.5%, and 35.9% in M04, respectively. Furthermore, the number of buds and branches treated with a mixture concentration on the the two clones increased by 90.7% and 21.7%, respectively. These data showed that the application of T. asperellum to cacao scions while grafting can decrease VSD incidence in side grafts and increase growth of grafts in addition to decreasing total number of stomata, increasing total area of opened stomata, and increasing number of buds and branches.

Health Benefits of Methylxanthines in Cacao and Chocolate

PubMed Central

Franco, Rafael; Oñatibia-Astibia, Ainhoa; Martínez-Pinilla, Eva

2013-01-01

One may wonder why methylxanthines are so abundant in beverages used by humans for centuries, or in cola-drinks that have been heavily consumed since their appearance. It is likely that humans have stuck to any brew containing compounds with psychoactive properties, resulting in a better daily life, i.e., more efficient thinking, exploring, hunting, etc., however, without the serious side effects of drugs of abuse. The physiological effects of methylxanthines have been known for a long time and they are mainly mediated by the so-called adenosine receptors. Caffeine and theobromine are the most abundant methylxanthines in cacao and their physiological effects are notable. Their health-promoting benefits are so remarkable that chocolate is explored as a functional food. The consequences of adenosine receptor blockade by natural compounds present in cacao/chocolate are here reviewed. Palatability and health benefits of methylxanthines, in general, and theobromine, in particular, have further contributed to sustain one of the most innocuous and pleasant habits: chocolate consumption. PMID:24145871

Molecular variability analysis of five new complete cacao swollen shoot virus genomic sequences.

PubMed

Muller, E; Sackey, S

2005-01-01

Cacao swollen shoot virus (CSSV), a member of the family Caulimovi-ridae, genus Badnavirus occurs in all the main cacao-growing areas of West Africa. We amplified, cloned and sequenced complete genomes of five new isolates, two originating from Togo and three originating from Ghana. The genome of these five newly sequenced isolates all contain the five putative open reading frames I, II, III, X and Y described for the first sequenced CSSV isolate, Agou1 originating from Togo. Their genomes have been aligned with the genome of Agou1. The nucleotide and amino acid sequence identities between isolates have been calculated and a phylogenetic analysis has been made including other pararetroviruses. Maximum nucleotide sequence variability between complete genomes of CSSV isolates was 29.4%. Geographical differentiation between isolates appears more important than differentiation between mild and severe isolates. ORF X differs greatly in size and sequence between the Togolese isolates Nyongbo2 and Agou1, and the four other isolates, its functional role is therefore clearly questionable.

Health benefits of methylxanthines in cacao and chocolate.

PubMed

Franco, Rafael; Oñatibia-Astibia, Ainhoa; Martínez-Pinilla, Eva

2013-10-18

One may wonder why methylxanthines are so abundant in beverages used by humans for centuries, or in cola-drinks that have been heavily consumed since their appearance. It is likely that humans have stuck to any brew containing compounds with psychoactive properties, resulting in a better daily life, i.e., more efficient thinking, exploring, hunting, etc., however, without the serious side effects of drugs of abuse. The physiological effects of methylxanthines have been known for a long time and they are mainly mediated by the so-called adenosine receptors. Caffeine and theobromine are the most abundant methylxanthines in cacao and their physiological effects are notable. Their health-promoting benefits are so remarkable that chocolate is explored as a functional food. The consequences of adenosine receptor blockade by natural compounds present in cacao/chocolate are here reviewed. Palatability and health benefits of methylxanthines, in general, and theobromine, in particular, have further contributed to sustain one of the most innocuous and pleasant habits: chocolate consumption.

Chemical speciation of cadmium: An approach to evaluate plant-available cadmium in Ecuadorian soils under cacao production.

PubMed

Chavez, E; He, Z L; Stoffella, P J; Mylavarapu, R S; Li, Y C; Baligar, V C

2016-05-01

Elevated concentration of cadmium (Cd) in cacao beans has raised serious concerns about the chocolate consumption on human health. Accumulation of Cd in cacao bean in southern Ecuador has been related to soil contamination. In this study, soil fractionation approach was used to identify available Cd pools in the soils and to correlate these Cd pools with bean Cd concentration and soil test indexes. The distribution of soil Cd fractions decreased in the order: oxidizable > acid-soluble > residual > reducible > water-soluble (+exchangeable). Oxidizable and acid-soluble fractions accounted for 59 and 68% of the total recoverable Cd for the 0-5 and 5-15 cm soil depth, respectively. Acid-soluble fraction was closely related to bean-Cd, with correlation coefficients (r) of 0.70 and 0.81 (P < 0.01) for the 0-5 and 5-15 cm soil depth, respectively. Acid-soluble Cd was significantly correlated with 0.01 M HCl- (r = 0.99, P < 0.01) or Mehlich 3- extractable Cd (r = 0.97, P < 0.01). These results indicate that acid-soluble Cd fraction is an important part of available Cd pool. Since approximately 60% of Cd in the cacao-growing soils is related to the acid-soluble fraction and bound to organic matter, remediation of the contaminated soils should consider to the dynamics of soil pH and organic matter content. Copyright © 2016 Elsevier Ltd. All rights reserved.

'Chocolate' silver nanoparticles: Synthesis, antibacterial activity and cytotoxicity.

PubMed

Chowdhury, Neelika Roy; MacGregor-Ramiasa, Melanie; Zilm, Peter; Majewski, Peter; Vasilev, Krasimir

2016-11-15

Silver nanoparticles (AgNPs) have emerged as a powerful weapon against antibiotic resistant microorganisms. However, most conventional AgNPs syntheses require the use of hazardous chemicals and generate toxic organic waste. Hence, in recent year's, plant derived and biomolecule based synthetics have has gained much attention. Cacao has been used for years for its medicinal benefits and contains a powerful reducing agent - oxalic acid. We hypothesized that, due to the presence of oxalic acid, cacao extract is capable of reducing silver nitrate (AgNO3) to produce AgNPs. In this study, AgNPs were synthesized by using natural cacao extract as a reducing and stabilizing agent. The reaction temperature, time and reactant molarity were varied to optimize the synthesis yield. UV-visible spectroscopy (UV-vis), dynamic light scattering (DLS) and transmission electron microscopy (TEM) characterization demonstrated that the synthesized AgNPs were spherical particles ranging in size from 35 to 42.5nm. The synthesized AgNPs showed significant antibacterial activity against clinically relevant pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis. Importantly, these green AgNPs are not cytotoxic to human dermal fibroblasts (HDFs) at concentrations below 32μg/ml. We conclude that cacao-based synthesis is a reproducible and sustainable method for the generation of stable antimicrobial silver nanoparticles with low cytotoxicity to human cells. The AgNPs synthesized in this work have promising properties for applications in the biomedical field. Copyright © 2016 Elsevier Inc. All rights reserved.

Apoplastic and intracellular plant sugars regulate developmental transitions in witches' broom disease of cacao.

PubMed

Barau, Joan; Grandis, Adriana; Carvalho, Vinicius Miessler de Andrade; Teixeira, Gleidson Silva; Zaparoli, Gustavo Henrique Alcalá; do Rio, Maria Carolina Scatolin; Rincones, Johana; Buckeridge, Marcos Silveira; Pereira, Gonçalo Amarante Guimarães

2015-03-01

Witches' broom disease (WBD) of cacao differs from other typical hemibiotrophic plant diseases by its unusually long biotrophic phase. Plant carbon sources have been proposed to regulate WBD developmental transitions; however, nothing is known about their availability at the plant-fungus interface, the apoplastic fluid of cacao. Data are provided supporting a role for the dynamics of soluble carbon in the apoplastic fluid in prompting the end of the biotrophic phase of infection. Carbon depletion and the consequent fungal sensing of starvation were identified as key signalling factors at the apoplast. MpNEP2, a fungal effector of host necrosis, was found to be up-regulated in an autophagic-like response to carbon starvation in vitro. In addition, the in vivo artificial manipulation of carbon availability in the apoplastic fluid considerably modulated both its expression and plant necrosis rate. Strikingly, infected cacao tissues accumulated intracellular hexoses, and showed stunted photosynthesis and the up-regulation of senescence markers immediately prior to the transition to the necrotrophic phase. These opposite findings of carbon depletion and accumulation in different host cell compartments are discussed within the frame of WBD development. A model is suggested to explain phase transition as a synergic outcome of fungal-related factors released upon sensing of extracellular carbon starvation, and an early senescence of infected tissues probably triggered by intracellular sugar accumulation. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

[Polyphenol content and antioxidant activity of several seeds and nuts].

PubMed

Padilla, F C; Rincón, A M; Bou-Rached, L

2008-09-01

Foods from plant origin not only provide human diet with certain antioxidant vitamins (C, E and beta-carotene), but also a complex mixture of polyphenols, with antioxidant activity. Numerous studies have been focused on the protective and preventing effect of this antioxidant activity on certain degenerative illnesses such as cardiovascular, cancer, and neurological diseases, cataracts and oxidative stress dysfunctions. The objective of this work was to evaluate total polyphenol content and antioxidant activity of several seeds, nuts, or grains such as Theobroma cacao, Canpsiandra comosa Benth (chiga), Sorghum bicolor, L. Moench, Melicoccus bijugatus (genip). Total polyphenol content was assessed by the Folin-Ciocalteau method and the antioxidant activity by the beta carotene/linoleate, reducing power, and the anti-radical activity methods. Results showed genip pericarp with the lowest polyphenol content (1.40 gGAE/100 g), and cacao beans with the highest (6.66 gGAE/100 g). Reducing power of cacao beans was also the highest and similar to the reducing power of 5.80 g ascorbic acid/100 g, followed by Campsiandra comosa. Moreover, Campsiandra comosa and cacao seeds presented an antioxidant activity comparable to that of the butylhydroxianisol, a synthetic antioxidant. The highest anti-radical activity was shown by Campsiandra comosa with an EC50 of 2.67 g/gDPPH. Total polyphenol content shows a good correlation with the antioxidant activity. Moreover, these seeds might have the same health beneficial effects attributed to other fruits and vegetables.

Combination of RNAseq and SNP nanofluidic array reveals the center of genetic diversity of cacao pathogen Moniliophthora roreri in the upper Magdalena Valley of Colombia and its clonality.

PubMed

Ali, Shahin S; Shao, Jonathan; Strem, Mary D; Phillips-Mora, Wilberth; Zhang, Dapeng; Meinhardt, Lyndel W; Bailey, Bryan A

2015-01-01

Moniliophthora roreri is the fungal pathogen that causes frosty pod rot (FPR) disease of Theobroma cacao L., the source of chocolate. FPR occurs in most of the cacao producing countries in the Western Hemisphere, causing yield losses up to 80%. Genetic diversity within the FPR pathogen population may allow the population to adapt to changing environmental conditions and adapt to enhanced resistance in the host plant. The present study developed single nucleotide polymorphism (SNP) markers from RNASeq results for 13 M. roreri isolates and validated the markers for their ability to reveal genetic diversity in an international M. roreri collection. The SNP resources reported herein represent the first study of RNA sequencing (RNASeq)-derived SNP validation in M. roreri and demonstrates the utility of RNASeq as an approach for de novo SNP identification in M. roreri. A total of 88 polymorphic SNPs were used to evaluate the genetic diversity of 172 M. roreri cacao isolates resulting in 37 distinct genotypes (including 14 synonymous groups). Absence of heterozygosity for the 88 SNP markers indicates reproduction in M. roreri is clonal and likely due to a homothallic life style. The upper Magdalena Valley of Colombia showed the highest levels of genetic diversity with 20 distinct genotypes of which 13 were limited to this region, and indicates this region as the possible center of origin for M. roreri.

Combination of RNAseq and SNP nanofluidic array reveals the center of genetic diversity of cacao pathogen Moniliophthora roreri in the upper Magdalena Valley of Colombia and its clonality

PubMed Central

Ali, Shahin S.; Shao, Jonathan; Strem, Mary D.; Phillips-Mora, Wilberth; Zhang, Dapeng; Meinhardt, Lyndel W.; Bailey, Bryan A.

2015-01-01

Moniliophthora roreri is the fungal pathogen that causes frosty pod rot (FPR) disease of Theobroma cacao L., the source of chocolate. FPR occurs in most of the cacao producing countries in the Western Hemisphere, causing yield losses up to 80%. Genetic diversity within the FPR pathogen population may allow the population to adapt to changing environmental conditions and adapt to enhanced resistance in the host plant. The present study developed single nucleotide polymorphism (SNP) markers from RNASeq results for 13 M. roreri isolates and validated the markers for their ability to reveal genetic diversity in an international M. roreri collection. The SNP resources reported herein represent the first study of RNA sequencing (RNASeq)-derived SNP validation in M. roreri and demonstrates the utility of RNASeq as an approach for de novo SNP identification in M. roreri. A total of 88 polymorphic SNPs were used to evaluate the genetic diversity of 172 M. roreri cacao isolates resulting in 37 distinct genotypes (including 14 synonymous groups). Absence of heterozygosity for the 88 SNP markers indicates reproduction in M. roreri is clonal and likely due to a homothallic life style. The upper Magdalena Valley of Colombia showed the highest levels of genetic diversity with 20 distinct genotypes of which 13 were limited to this region, and indicates this region as the possible center of origin for M. roreri. PMID:26379633

Land use change effects on trace gas fluxes in the forest margins of Central Sulawesi, Indonesia

NASA Astrophysics Data System (ADS)

Veldkamp, Edzo; Purbopuspito, Joko; Corre, Marife D.; Brumme, Rainer; Murdiyarso, Daniel

2008-06-01

Land use changes and land use intensification are considered important processes contributing to the increasing concentrations of the greenhouse gases nitrous oxide (N2O) and methane (CH4) and of nitric oxide (NO), a precursor of ozone. Studies on the effects of land use changes and land use intensification on soil trace gas emissions were mostly conducted in Latin America and only very few in Asia. Here we present results from Central Sulawesi where profound changes in land use and cultivation practices take place: traditional agricultural practices like shifting cultivation and slash-and-burn agriculture are replaced by permanent cultivation systems and introduction of income-generating cash crops like cacao. Our results showed that N2O emissions were higher from cacao agroforestry (35 ± 10 μg N m-2 h-1) than maize (9 ± 2 μg N m-2 h-1), whereas intermediate rates were observed from secondary forests (25 ± 11 μg N m-2 h-1). NO emissions did not differ among land use systems, ranging from 12 ± 2 μg N m-2 h-1 for cacao agroforestry and secondary forest to 18 ± 2 μg N m-2 h-1 for maize. CH4 uptake was higher for maize (-30 ± 4 μg C m-2 h-1) than cacao agroforestry (-18 ± 2 μg C m-2 h-1) and intermediate rates were measured from secondary forests (-25 ± 4 μg C m-2 h-1). Combining these data with results from other studies in this area, we present chronosequence effects of land use change on trace gas emissions from natural forest, through maize cultivation, to cacao agroforestry (with or without fertilizer). Compared to the original forests, this typical land use change in the study area clearly led to higher N2O emissions and lower CH4 uptake with age of cacao agroforestry systems. We conclude that this common land use sequence in the area combined with the increasing use of fertilizer will strongly increase soil trace gas emissions. We suggest that the future hot spot regions of high N2O (and to a lesser extend NO) emissions in the tropics are those areas where climatic and edaphic conditions allow for intensive agriculture. This scenario is probably preferable over the alternative of agriculture extensification, which would imply a dramatic increase in deforestation rates with accompanying CO2 emissions.

A fast algorithm for computer aided collimation gamma camera (CACAO)

NASA Astrophysics Data System (ADS)

Jeanguillaume, C.; Begot, S.; Quartuccio, M.; Douiri, A.; Franck, D.; Pihet, P.; Ballongue, P.

2000-08-01

The computer aided collimation gamma camera is aimed at breaking down the resolution sensitivity trade-off of the conventional parallel hole collimator. It uses larger and longer holes, having an added linear movement at the acquisition sequence. A dedicated algorithm including shift and sum, deconvolution, parabolic filtering and rotation is described. Examples of reconstruction are given. This work shows that a simple and fast algorithm, based on a diagonal dominant approximation of the problem can be derived. Its gives a practical solution to the CACAO reconstruction problem.

Fading positive effect of biochar on crop yield and soil acidity during five growth seasons in an Indonesian Ultisol.

PubMed

Cornelissen, Gerard; Jubaedah; Nurida, Neneng L; Hale, Sarah E; Martinsen, Vegard; Silvani, Ludovica; Mulder, Jan

2018-09-01

Low fertility limits crop production on acidic soils dominating much of the humid tropics. Biochar may be used as a soil enhancer, but little consensus exists on its effect on crop yield. Here we use a controlled, replicated and long-term field study in Sumatra, Indonesia, to investigate the longevity and mechanism of the effects of two contrasting biochars (produced from rice husk and cacao shell, and applied at dosages of 5 and 15tha -1 ) on maize production in a highly acidic Ultisol (pH KCl 3.6). Compared to rice husk biochar, cacao shell biochar exhibited a higher pH (9.8 vs. 8.4), CEC (197 vs. 20cmol c kg -1 ) and acid neutralizing capacity (217 vs. 45cmol c kg -1 ) and thus had a greater liming potential. Crop yield effects of cacao shell biochar (15tha -1 ) were also much stronger than those of rice husk biochar, and could be related to more favorable Ca/Al ratios in response to cacao shell biochar (1.0 to 1.5) compared to rice husk biochar (0.3 to 0.6) and nonamended plots (0.15 to 0.6). The maize yield obtained with the cacao shell biochar peaked in season 2, continued to have a good effect in seasons 3-4, and faded in season 5. The yield effect of the rice husk biochar was less pronounced and already faded from season 2 onwards. Crop yields were correlated with the pH-related parameters Ca/Al ratio, base saturation and exchangeable K. The positive effects of cocoa shell biochar on crop yield in this Ultisol were at least in part related to alleviation of soil acidity. The fading effectiveness after multiple growth seasons, possibly due to leaching of the biochar-associated alkalinity, indicates that 15tha -1 of cocoa shell biochar needs to be applied approximately every third season in order to maintain positive effects on yield. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Sodium-potassium synergism in Theobroma cacao: stimulation of photosynthesis, water-use efficiency and mineral nutrition.

PubMed

Gattward, James N; Almeida, Alex-Alan F; Souza, José O; Gomes, Fábio P; Kronzucker, Herbert J

2012-11-01

In ecological setting, sodium (Na(+)) can be beneficial or toxic, depending on plant species and the Na(+) level in the soil. While its effects are more frequently studied at high saline levels, Na(+) has also been shown to be of potential benefit to some species at lower levels of supply, especially in C4 species. Here, clonal plants of the major tropical C3 crop Theobroma cacao (cacao) were grown in soil where potassium (K(+)) was partially replaced (at six levels, up to 50% replacement) by Na(+), at two concentrations (2.5 and 4.0 mmol(c) dm(-3)). At both concentrations, net photosynthesis per unit leaf area (A) increased more than twofold with increasing substitution of K(+) by Na(+). Concomitantly, instantaneous (A/E) and intrinsic (A/g(s)) water-use efficiency (WUE) more than doubled. Stomatal conductance (g(s)) and transpiration rate (E) exhibited a decline at 2.5 mmol dm(-3), but remained unchanged at 4 mmol dm(-3). Leaf nitrogen content was not impacted by Na(+) supplementation, whereas sulfur (S), calcium (Ca(2+)), magnesium (Mg(2+)) and zinc (Zn(2+)) contents were maximized at 2.5 mmol dm(-3) and intermediate (30-40%) replacement levels. Leaf K(+) did not decline significantly. In contrast, leaf Na(+) content increased steadily. The resultant elevated Na(+)/K(+) ratios in tissue correlated with increased, not decreased, plant performance. The results show that Na(+) can partially replace K(+) in the nutrition of clonal cacao, with significant beneficial effects on photosynthesis, WUE and mineral nutrition in this major perennial C3 crop. Copyright © Physiologia Plantarum 2012.

Independent Origins of Yeast Associated with Coffee and Cacao Fermentation.

PubMed

Ludlow, Catherine L; Cromie, Gareth A; Garmendia-Torres, Cecilia; Sirr, Amy; Hays, Michelle; Field, Colburn; Jeffery, Eric W; Fay, Justin C; Dudley, Aimée M

2016-04-04

Modern transportation networks have facilitated the migration and mingling of previously isolated populations of plants, animals, and insects. Human activities can also influence the global distribution of microorganisms. The best-understood example is yeasts associated with winemaking. Humans began making wine in the Middle East over 9,000 years ago [1, 2]. Selecting favorable fermentation products created specialized strains of Saccharomyces cerevisiae [3, 4] that were transported along with grapevines. Today, S. cerevisiae strains residing in vineyards around the world are genetically similar, and their population structure suggests a common origin that followed the path of human migration [3-7]. Like wine, coffee and cacao depend on microbial fermentation [8, 9] and have been globally dispersed by humans. Theobroma cacao originated in the Amazon and Orinoco basins of Colombia and Venezuela [10], was cultivated in Central America by Mesoamerican peoples, and was introduced to Europeans by Hernán Cortés in 1530 [11]. Coffea, native to Ethiopia, was disseminated by Arab traders throughout the Middle East and North Africa in the 6(th) century and was introduced to European consumers in the 17(th) century [12]. Here, we tested whether the yeasts associated with coffee and cacao are genetically similar, crop-specific populations or genetically diverse, geography-specific populations. Our results uncovered populations that, while defined by niche and geography, also bear signatures of admixture between major populations in events independent of the transport of the plants. Thus, human-associated fermentation and migration may have affected the distribution of yeast involved in the production of coffee and chocolate. Copyright © 2016 Elsevier Ltd. All rights reserved.

Phytophthora megakarya and Phytophthora palmivora, Closely Related Causal Agents of Cacao Black Pod Rot, Underwent Increases in Genome Sizes and Gene Numbers by Different Mechanisms

PubMed Central

Ali, Shahin S.; Shao, Jonathan; Lary, David J.; Kronmiller, Brent A.; Shen, Danyu; Strem, Mary D.; Amoako-Attah, Ishmael; Akrofi, Andrew Yaw; Begoude, B.A. Didier; ten Hoopen, G. Martijn; Coulibaly, Klotioloma; Kebe, Boubacar Ismaël; Melnick, Rachel L.; Guiltinan, Mark J.; Tyler, Brett M.; Meinhardt, Lyndel W.

2017-01-01

Phytophthora megakarya (Pmeg) and Phytophthora palmivora (Ppal) are closely related species causing cacao black pod rot. Although Ppal is a cosmopolitan pathogen, cacao is the only known host of economic importance for Pmeg. Pmeg is more virulent on cacao than Ppal. We sequenced and compared the Pmeg and Ppal genomes and identified virulence-related putative gene models (PGeneM) that may be responsible for their differences in host specificities and virulence. Pmeg and Ppal have estimated genome sizes of 126.88 and 151.23 Mb and PGeneM numbers of 42,036 and 44,327, respectively. The evolutionary histories of Pmeg and Ppal appear quite different. Postspeciation, Ppal underwent whole-genome duplication whereas Pmeg has undergone selective increases in PGeneM numbers, likely through accelerated transposable element-driven duplications. Many PGeneMs in both species failed to match transcripts and may represent pseudogenes or cryptic genetic reservoirs. Pmeg appears to have amplified specific gene families, some of which are virulence-related. Analysis of mycelium, zoospore, and in planta transcriptome expression profiles using neural network self-organizing map analysis generated 24 multivariate and nonlinear self-organizing map classes. Many members of the RxLR, necrosis-inducing phytophthora protein, and pectinase genes families were specifically induced in planta. Pmeg displays a diverse virulence-related gene complement similar in size to and potentially of greater diversity than Ppal but it remains likely that the specific functions of the genes determine each species’ unique characteristics as pathogens. PMID:28186564

Recombinant β-1,3-1,4-glucanase from Theobroma cacao impairs Moniliophthora perniciosa mycelial growth.

PubMed

Britto, Dahyana Santos; Pirovani, Carlos Priminho; Andrade, Bruno Silva; Dos Santos, Tassiara Pereira; Pungartnik, Cristina; Cascardo, Júlio Cezar M; Micheli, Fabienne; Gesteira, Abelmon S

2013-09-01

In this work, we identified a gene from Theobroma cacao L. genome and cDNA libraries, named TcGlu2, that encodes a β-1,3-1,4-glucanase. The TcGlu2 ORF was 720 bp in length and encoded a polypeptide of 239 amino acids with a molecular mass of 25.58 kDa. TcGlu2 contains a conserved domain characteristic of β-1,3-1,4-glucanases and presented high protein identity with β-1,3-1,4-glucanases from other plant species. Molecular modeling of TcGlu2 showed an active site of 13 amino acids typical of glucanase with β-1,3 and 1,4 action mode. The recombinant cDNA TcGlu2 obtained by heterologous expression in Escherichia coli and whose sequence was confirmed by mass spectrometry, has a molecular mass of about 22 kDa (with His-Tag) and showed antifungal activity against the fungus Moniliophthora perniciosa, causal agent of the witches' broom disease in cacao. The integrity of the hyphae membranes of M. perniciosa, incubated with protein TcGlu2, was analyzed with propidium iodide. After 1 h of incubation, a strong fluorescence emitted by the hyphae indicating the hydrolysis of the membrane by TcGlu2, was observed. To our knowledge, this is the first study of a cacao β-1,3-1,4-glucanase expression in heterologous system and the first analysis showing the antifungal activity of a β-1,3-1,4-glucanase, in particular against M. perniciosa.

Preparative separation of cacao bean procyanidins by high-speed counter-current chromatography.

PubMed

Li, Lingxi; Zhang, Shuting; Cui, Yan; Li, Yuanyuan; Luo, Lanxin; Zhou, Peiyu; Sun, Baoshan

2016-11-15

In this work, an efficient method for preparative separation of procyanidins from raw cacao bean extract by high-speed counter-current chromatography (HSCCC) was developed. Under the optimized solvent system of n-hexane-ethyl acetate-water (1:50:50, v/v/v) with a combination of head-tail and tail-head elution modes, various procyanidins fractions with different polymerization degrees were successfully separated. UPLC, QTOF-MS and 1 H NMR analysis verified that these fractions contained monomer up to pentamer respectively. Dimeric procyanidin B2 (purity>86%) could be isolated by HSCCC in a single run. Other individual procyanidins in these fractions could be further isolated and purified by preparative HPLC. The developed HSCCC together with preparative HPLC techniques appeared to be a useful tool for large preparation of different procyanidins from cacao beans. Furthermore, by antioxidant activity assays, it was proved that both fractions and individual procyanidins possessed greater antioxidant activities compared to standard trolox. The antioxidant activities of procyanidins increase as the increase of their polymerization degree. Copyright © 2016 Elsevier B.V. All rights reserved.

Why are there few seedlings beneath the myrmecophyte Triplaris americana?

NASA Astrophysics Data System (ADS)

Larrea-Alcázar, Daniel M.; Simonetti, Javier A.

2007-07-01

We compared the relative importance of chemical alellopathy, pruning behaviour of resident ants and other non-related agents to ant-plant mutualism for seedling establishment beneath Triplaris americana L. (Polygonaceae), a myrmecophyte plant. We also included a preliminary analysis of effects of fragmentation on these ecological processes. Seeds and seedlings of Theobroma cacao L. (Sterculiaceae) were used as the target species in all experiments. Leaf-tissue extracts of the myrmecophyte plant did not inhibit germination of cacao seeds. Resident Pseudomyrmex triplarinus Weddell (Pseudomyrmecinae) ants did not remove seeds under the canopy of their host plants. The main seed consumer was the leaf-cutting ant Atta sexdens L. (Myrmicinae). Leaves of cacao seedlings were partially or totally pruned by Pseudomyrmex ants mainly in forest fragments studied. We offer evidence pointing to the possibility that the absence of seedlings beneath Triplaris may result from effects of both ant species. We discuss the benefits of pruning behaviour for the resident ant colony and the effects of ant-ant interactions on seedling establishment beneath this ant-plant system.

Profiles of phenolic compounds and purine alkaloids during the development of seeds of Theobroma cacao cv. Trinitario.

PubMed

Pereira-Caro, Gema; Borges, Gina; Nagai, Chifumi; Jackson, Mel C; Yokota, Takao; Crozier, Alan; Ashihara, Hiroshi

2013-01-16

Changes occurring in phenolic compounds and purine alkaloids, during the growth of seeds of cacao (Theobroma cacao) cv. Trinitario, were investigated using HPLC-MS/MS. Extracts of seeds with a fresh weight of 125, 700, 1550, and 2050 mg (stages 1-4, respectively) were analyzed. The phenolic compounds present in highest concentrations in developing and mature seeds (stages 3 and 4) were flavonols and flavan-3-ols. Flavan-3-ols existed as monomers of epicatechin and catechin and as procyanidins. Type B procyanidins were major components and varied from dimers to pentadecamer. Two anthocyanins, cyanidin-3-O-arabinoside and cyanidin-3-O-galactoside, along with the N-phenylpropernoyl-l-amino acids, N-caffeoyl-l-aspartate, N-coumaroyl-l-aspartate, N-coumaroyl-3-hydroxytyrosine (clovamide), and N-coumaroyltyrosine (deoxyclovamide), and the purine alkaloids theobromine and caffeine, were present in stage 3 and 4 seeds. Other purine alkaloids, such as theophylline and additional methylxanthines, did not occur in detectable quantities. Flavan-3-ols were the only components to accumulate in detectable quantities in young seeds at developmental stages 1 and 2.

Isolation of dimeric, trimeric, tetrameric and pentameric procyanidins from unroasted cocoa beans (Theobroma cacao L.) using countercurrent chromatography.

PubMed

Esatbeyoglu, Tuba; Wray, Victor; Winterhalter, Peter

2015-07-15

The main procyanidins, including dimeric B2 and B5, trimeric C1, tetrameric and pentameric procyanidins, were isolated from unroasted cocoa beans (Theobroma cacao L.) using various techniques of countercurrent chromatography, such as high-speed countercurrent chromatography (HSCCC), low-speed rotary countercurrent chromatography (LSRCCC) and spiral-coil LSRCCC. Furthermore, dimeric procyanidins B1 and B7, which are not present naturally in the analysed cocoa beans, were obtained after semisynthesis of cocoa bean polymers with (+)-catechin as nucleophile and separated by countercurrent chromatography. In this way, the isolation of dimeric procyanidin B1 in considerable amounts (500mg, purity>97%) was possible in a single run. This is the first report concerning the isolation and semisynthesis of dimeric to pentameric procyanidins from T. cacao by countercurrent chromatography. Additionally, the chemical structures of tetrameric (cinnamtannin A2) and pentameric procyanidins (cinnamtannin A3) were elucidated on the basis of (1)H NMR spectroscopy. Interflavanoid linkage was determined by NOE-correlations, for the first time. Copyright © 2015 Elsevier Ltd. All rights reserved.

Simultaneous decontamination of cross-polluted soils with heavy metals and PCBs using a nano-metallic Ca/CaO dispersion mixture.

PubMed

Mallampati, Srinivasa Reddy; Mitoma, Yoshiharu; Okuda, Tetsuji; Sakita, Shogo; Simion, Cristian

2014-01-01

In the present work, we investigated the use of nano-metallic calcium (Ca) and calcium oxide (CaO) dispersion mixture for the simultaneous remediation of contaminated soils with both heavy metals (As, Cd, Cr, and Pb) and polychlorinated biphenyls (PCBs). Regardless of soil moisture content, nano-metallic Ca/CaO dispersion mixture achieved about 95-99% of heavy metal immobilization by a simple grinding process. During the same treatment, reasonable PCB hydrodechlorination efficiencies were obtained (up to 97%), though higher hydrodechlorination efficiency by preliminary drying of soil was observed.

Combining-ability for cacao (Theobroma cacao L.) yield components under southern Bahia conditions.

PubMed

Dias, L A; Kageyama, P Y

1995-03-01

The objective of this study was to assess five cacao cultivars (selfs) and 20 hybrids with regard to their general-and specific-combining ability for yield components using method 1, model I, of the diallel analysis system. The selfings and the hybrids were obtained through controlled crossings, tested in the field in a random block design with four replications and plots containing 16 plants. The experiment was set up in the Centro de Pesquisas do Cacau, in Itabuna, Bahia, Brasil, in 1975. The characteristics studied were: the number of healthy and collected fruits per plant (NHFP and NCFP), the weight of humid seeds per plant and per fruit (WHSP and WHSF), and the percentage of diseased fruits per plant (PDFP), for 5 years (1986-1990). The F-test values, highly significant for general combining ability (GCA) and specific combining ability (SCA), demonstrated the existence of variability for both effects. However, the effects of SCA were greater than those of GCA, when compared in terms of the average squared effects. This condition held for the characteristics NHFP, NCFP and WHSP, which shows the relative importance of the non-additive genetic effects over the additive effects. The reciprocal effects did not show significance. Breeding methods which explore the additive portion of genetic variance should be employed for obtaining higher-yielding cacao and high seed weight. For this, the segregant populations should involve cultivars CEPEC 1, SIAL 169 and ICS 1. Combinations involving the cultivar ICS 1 presented the most favorable results for the characteristics WHSP and WHSF, where the hybrid SIAL 169 x ICS 1 and its reciprocal were outstanding.

First Microsatellite Markers Developed from Cupuassu ESTs: Application in Diversity Analysis and Cross-Species Transferability to Cacao.

PubMed

Ferraz Dos Santos, Lucas; Moreira Fregapani, Roberta; Falcão, Loeni Ludke; Togawa, Roberto Coiti; Costa, Marcos Mota do Carmo; Lopes, Uilson Vanderlei; Peres Gramacho, Karina; Alves, Rafael Moyses; Micheli, Fabienne; Marcellino, Lucilia Helena

2016-01-01

The cupuassu tree (Theobroma grandiflorum) (Willd. ex Spreng.) Schum. is a fruitful species from the Amazon with great economical potential, due to the multiple uses of its fruit´s pulp and seeds in the food and cosmetic industries, including the production of cupulate, an alternative to chocolate. In order to support the cupuassu breeding program and to select plants presenting both pulp/seed quality and fungal disease resistance, SSRs from Next Generation Sequencing ESTs were obtained and used in diversity analysis. From 8,330 ESTs, 1,517 contained one or more SSRs (1,899 SSRs identified). The most abundant motifs identified in the EST-SSRs were hepta- and trinucleotides, and they were found with a minimum and maximum of 2 and 19 repeats, respectively. From the 1,517 ESTs containing SSRs, 70 ESTs were selected based on their functional annotation, focusing on pulp and seed quality, as well as resistance to pathogens. The 70 ESTs selected contained 77 SSRs, and among which, 11 were polymorphic in cupuassu genotypes. These EST-SSRs were able to discriminate the cupuassu genotype in relation to resistance/susceptibility to witches' broom disease, as well as to pulp quality (SST/ATT values). Finally, we showed that these markers were transferable to cacao genotypes, and that genome availability might be used as a predictive tool for polymorphism detection and primer design useful for both Theobroma species. To our knowledge, this is the first report involving EST-SSRs from cupuassu and is also a pioneer in the analysis of marker transferability from cupuassu to cacao. Moreover, these markers might contribute to develop or saturate the cupuassu and cacao genetic maps, respectively.

Molecular, Biochemical and Ultrastructural Changes Induced by Pb Toxicity in Seedlings of Theobroma cacao L.

PubMed

Reis, Graciele Santos Monteiro; de Almeida, Alex-Alan Furtado; de Almeida, Nicolle Moreira; de Castro, Andressa Vieira; Mangabeira, Pedro Antonio Oliveira; Pirovani, Carlos Priminho

2015-01-01

Pb is a metal which is highly toxic to plants and animals, including humans. High concentrations of Pb have been observed in beans of T. cacao, as well as in its products. In this work, we evaluated the molecular, biochemical, and ultrastructural alterations in mature leaves and primary roots of seedlings of two progenies of T. cacao, obtained from seed germination in different concentrations of Pb (0, 0.05, 0.1, 0.2, 0.4, 0.8 g L(-1)), in the form of Pb(NO3)2. The progenies resulted from self-fertilization of Catongo and a cross of CCN-10 x SCA-6. The Pb, supplied via seminal, caused alterations in the ultrastructures of the mesophyll cells and in the amount of starch grains in the chloroplasts. The dosage of substances reactive to thiobarbituric acid showed that Pb induced lipid peroxidation. The activity of guaiacol peroxidases and the expression of genes associated to synthetase of phytochelatin, SODcyt and PER increased in response to Pb. In addition, there was alteration in the expression of stress-related proteins. The progeny of CCN-10 x SCA-6 was more tolerant to Pb stress when compared to Catongo, since: (i) it accumulated more Pb in the roots, preventing its translocation to the shoot; (ii) it presented higher activity of peroxidases in the roots, which are enzymes involved in the elimination of excess of reactive oxygen species; and (iii) increased expression of the gene in the phytochelatin biosynthesis route. The results of the proteomic analysis were of paramount importance to differentiate the defense mechanisms used by both progenies of T. cacao.

First Microsatellite Markers Developed from Cupuassu ESTs: Application in Diversity Analysis and Cross-Species Transferability to Cacao

PubMed Central

Ferraz dos Santos, Lucas; Moreira Fregapani, Roberta; Falcão, Loeni Ludke; Togawa, Roberto Coiti; Costa, Marcos Mota do Carmo; Lopes, Uilson Vanderlei; Peres Gramacho, Karina; Alves, Rafael Moyses

2016-01-01

The cupuassu tree (Theobroma grandiflorum) (Willd. ex Spreng.) Schum. is a fruitful species from the Amazon with great economical potential, due to the multiple uses of its fruit´s pulp and seeds in the food and cosmetic industries, including the production of cupulate, an alternative to chocolate. In order to support the cupuassu breeding program and to select plants presenting both pulp/seed quality and fungal disease resistance, SSRs from Next Generation Sequencing ESTs were obtained and used in diversity analysis. From 8,330 ESTs, 1,517 contained one or more SSRs (1,899 SSRs identified). The most abundant motifs identified in the EST-SSRs were hepta- and trinucleotides, and they were found with a minimum and maximum of 2 and 19 repeats, respectively. From the 1,517 ESTs containing SSRs, 70 ESTs were selected based on their functional annotation, focusing on pulp and seed quality, as well as resistance to pathogens. The 70 ESTs selected contained 77 SSRs, and among which, 11 were polymorphic in cupuassu genotypes. These EST-SSRs were able to discriminate the cupuassu genotype in relation to resistance/susceptibility to witches’ broom disease, as well as to pulp quality (SST/ATT values). Finally, we showed that these markers were transferable to cacao genotypes, and that genome availability might be used as a predictive tool for polymorphism detection and primer design useful for both Theobroma species. To our knowledge, this is the first report involving EST-SSRs from cupuassu and is also a pioneer in the analysis of marker transferability from cupuassu to cacao. Moreover, these markers might contribute to develop or saturate the cupuassu and cacao genetic maps, respectively. PMID:26949967

Molecular cloning and expression analysis of the sucrose transporter gene family from Theobroma cacao L.

PubMed

Li, Fupeng; Wu, Baoduo; Qin, Xiaowei; Yan, Lin; Hao, Chaoyun; Tan, Lehe; Lai, Jianxiong

2014-08-10

In this study, we performed cloning and expression analysis of six putative sucrose transporter genes, designated TcSUT1, TcSUT2, TcSUT3, TcSUT4, TcSUT5 and TcSUT6, from the cacao genotype 'TAS-R8'. The combination of cDNA and genomic DNA sequences revealed that the cacao SUT genes contained exon numbers ranging from 1 to 14. The average molecular mass of all six deduced proteins was approximately 56 kDa (range 52 to 66 kDa). All six proteins were predicted to exhibit typical features of sucrose transporters with 12 trans-membrane spanning domains. Phylogenetic analysis revealed that TcSUT2 and TcSUT4 belonged to Group 2 SUT and Group 4 SUT, respectively, and the other TcSUT proteins were belonging to Group 1 SUT. Real-time PCR was conducted to investigate the expression pattern of each member of the SUT family in cacao. Our experiment showed that TcSUT1 was expressed dominantly in pods and that, TcSUT3 and TcSUT4 were highly expressed in both pods and in bark with phloem. Within pods, TcSUT1 and TcSUT4 were expressed more in the seed coat and seed from the pod enlargement stage to the ripening stage. TcSUT5 expression sharply increased to its highest expression level in the seed coat during the ripening stage. Expression pattern analysis indicated that TcSUT genes may be associated with photoassimilate transport into developing seeds and may, therefore, have an impact on seed production. Copyright © 2014 Elsevier B.V. All rights reserved.

Molecular, Biochemical and Ultrastructural Changes Induced by Pb Toxicity in Seedlings of Theobroma cacao L.

PubMed Central

Reis, Graciele Santos Monteiro; de Almeida, Alex-Alan Furtado; de Almeida, Nicolle Moreira; de Castro, Andressa Vieira; Mangabeira, Pedro Antonio Oliveira; Pirovani, Carlos Priminho

2015-01-01

Pb is a metal which is highly toxic to plants and animals, including humans. High concentrations of Pb have been observed in beans of T. cacao, as well as in its products. In this work, we evaluated the molecular, biochemical, and ultrastructural alterations in mature leaves and primary roots of seedlings of two progenies of T. cacao, obtained from seed germination in different concentrations of Pb (0, 0.05, 0.1, 0.2, 0.4, 0.8 g L-1), in the form of Pb(NO3)2. The progenies resulted from self-fertilization of Catongo and a cross of CCN-10 x SCA-6. The Pb, supplied via seminal, caused alterations in the ultrastructures of the mesophyll cells and in the amount of starch grains in the chloroplasts. The dosage of substances reactive to thiobarbituric acid showed that Pb induced lipid peroxidation. The activity of guaiacol peroxidases and the expression of genes associated to synthetase of phytochelatin, SODcyt and PER increased in response to Pb. In addition, there was alteration in the expression of stress-related proteins. The progeny of CCN-10 x SCA-6 was more tolerant to Pb stress when compared to Catongo, since: (i) it accumulated more Pb in the roots, preventing its translocation to the shoot; (ii) it presented higher activity of peroxidases in the roots, which are enzymes involved in the elimination of excess of reactive oxygen species; and (iii) increased expression of the gene in the phytochelatin biosynthesis route. The results of the proteomic analysis were of paramount importance to differentiate the defense mechanisms used by both progenies of T. cacao. PMID:26146994

Impact of fermentation, drying, roasting and Dutch processing on flavan-3-ol stereochemistry in cacao beans and cocoa ingredients

PubMed Central

2011-01-01

This paper reports a systematic study of the level of flavan-3-ol monomers during typical processing steps as cacao beans are dried, fermented and roasted and the results of Dutch-processing. Methods have been used that resolve the stereoisomers of epicatechin and catechin. In beans harvested from unripe and ripe cacao pods, we find only (-)-epicatechin and (+)-catechin with (-)-epicatechin being by far the predominant isomer. When beans are fermented there is a large loss of both (-)-epicatechin and (+)-catechin, but also the formation of (-)-catechin. We hypothesize that the heat of fermentation may, in part, be responsible for the formation of this enantiomer. When beans are progressively roasted at conditions described as low, medium and high roast conditions, there is a progressive loss of (-)-epicatechin and (+)-catechin and an increase in (-)-catechin with the higher roast levels. When natural and Dutch-processed cacao powders are analyzed, there is progressive loss of both (-)-epicatechin and (+)-catechin with lesser losses of (-)-catechin. We thus observe that in even lightly Dutch-processed powder, the level of (-)-catechin exceeds the level of (-)-epicatechin. The results indicate that much of the increase in the level of (-)-catechin observed during various processing steps may be the result of heat-related epimerization from (-)-epicatechin. These results are discussed with reference to the reported preferred order of absorption of (-)-epicatechin > (+)-catechin > (-)-catechin. These results are also discussed with respect to the balance that must be struck between the beneficial impact of fermentation and roasting on chocolate flavor and the healthful benefits of chocolate and cocoa powder that result in part from the flavan-3-ol monomers. PMID:21917164

The cacao Criollo genome v2.0: an improved version of the genome for genetic and functional genomic studies.

PubMed

Argout, X; Martin, G; Droc, G; Fouet, O; Labadie, K; Rivals, E; Aury, J M; Lanaud, C

2017-09-15

Theobroma cacao L., native to the Amazonian basin of South America, is an economically important fruit tree crop for tropical countries as a source of chocolate. The first draft genome of the species, from a Criollo cultivar, was published in 2011. Although a useful resource, some improvements are possible, including identifying misassemblies, reducing the number of scaffolds and gaps, and anchoring un-anchored sequences to the 10 chromosomes. We used a NGS-based approach to significantly improve the assembly of the Belizian Criollo B97-61/B2 genome. We combined four Illumina large insert size mate paired libraries with 52x of Pacific Biosciences long reads to correct misassembled regions and reduced the number of scaffolds. We then used genotyping by sequencing (GBS) methods to increase the proportion of the assembly anchored to chromosomes. The scaffold number decreased from 4,792 in assembly V1 to 554 in V2 while the scaffold N50 size has increased from 0.47 Mb in V1 to 6.5 Mb in V2. A total of 96.7% of the assembly was anchored to the 10 chromosomes compared to 66.8% in the previous version. Unknown sites (Ns) were reduced from 10.8% to 5.7%. In addition, we updated the functional annotations and performed a new RefSeq structural annotation based on RNAseq evidence. Theobroma cacao Criollo genome version 2 will be a valuable resource for the investigation of complex traits at the genomic level and for future comparative genomics and genetics studies in cacao tree. New functional tools and annotations are available on the Cocoa Genome Hub ( http://cocoa-genome-hub.southgreen.fr ).

Impact of fermentation, drying, roasting and Dutch processing on flavan-3-ol stereochemistry in cacao beans and cocoa ingredients.

PubMed

Hurst, W Jeffrey; Krake, Susann H; Bergmeier, Stephen C; Payne, Mark J; Miller, Kenneth B; Stuart, David A

2011-09-14

This paper reports a systematic study of the level of flavan-3-ol monomers during typical processing steps as cacao beans are dried, fermented and roasted and the results of Dutch-processing. Methods have been used that resolve the stereoisomers of epicatechin and catechin. In beans harvested from unripe and ripe cacao pods, we find only (-)-epicatechin and (+)-catechin with (-)-epicatechin being by far the predominant isomer. When beans are fermented there is a large loss of both (-)-epicatechin and (+)-catechin, but also the formation of (-)-catechin. We hypothesize that the heat of fermentation may, in part, be responsible for the formation of this enantiomer. When beans are progressively roasted at conditions described as low, medium and high roast conditions, there is a progressive loss of (-)-epicatechin and (+)-catechin and an increase in (-)-catechin with the higher roast levels. When natural and Dutch-processed cacao powders are analyzed, there is progressive loss of both (-)-epicatechin and (+)-catechin with lesser losses of (-)-catechin. We thus observe that in even lightly Dutch-processed powder, the level of (-)-catechin exceeds the level of (-)-epicatechin. The results indicate that much of the increase in the level of (-)-catechin observed during various processing steps may be the result of heat-related epimerization from (-)-epicatechin. These results are discussed with reference to the reported preferred order of absorption of (-)-epicatechin > (+)-catechin > (-)-catechin. These results are also discussed with respect to the balance that must be struck between the beneficial impact of fermentation and roasting on chocolate flavor and the healthful benefits of chocolate and cocoa powder that result in part from the flavan-3-ol monomers.

Antibacterial activity of alimentary plants against Staphylococcus aureus growth.

PubMed

Pérez, C; Anesini, C

1994-01-01

Alimentary plants were screened for antibacterial activity against a penicillin G resistant strain of Staphylococcus aureus. Twenty-five samples of plant material corresponding to 21 species from 13 families were used. Both aqueous and ethanol extracts were obtained from them. Antibacterial activity was determined by the agar-well diffusion method, using cephazolin as a standard antibiotic. Seventeen ethanol extracts were found active. Eugenia caryophyllata (clavo de olor*) flowers, Myristica fragans (nuez moscada*) seeds, Theobroma cacao (cacao*) seed bark, Triticum sp (trigo*) fruit, Zea mays (maíz*) fruit and Piper nigrum (pimienta*) ripe fruit produced some of the more active extracts (* = Argentine vulgar names).

1H NMR study of fermented cocoa (Theobroma cacao L.) beans.

PubMed

Caligiani, Augusta; Acquotti, Domenico; Cirlini, Martina; Palla, Gerardo

2010-12-08

This study reports for the first time the metabolic profile of cocoa (Theobroma cacao L.) beans using the (1)H NMR technique applied to polar extracts of fermented cocoa beans. The simultaneous detection and quantification of amino acids, polyalcohols, organic acids, sugars, methylxanthines, catechins, and phenols were obtained by assigning the major signals of the spectra for different varieties of cocoa beans (Forastero, Criollo, and Trinitario) from different countries (Ecuador, Ghana, Grenada, and Trinidad). The data set obtained, representative of all classes of soluble compounds of cocoa, was useful to characterize the fermented cocoa beans as a function of the variety and geographic origin.

Utilization of Cacao Pod Husk Silage as Cattle Ration Mixture at Taluditi, Pohuwatu Regency, Gorontalo Province

NASA Astrophysics Data System (ADS)

Hidayat, M.; Pratama, H. Y.; Martono, E.

2018-02-01

Cacao plantation produces cacao pod husks (CPHs) by-products during the harvest period. This research aimed to make benefits the CPH, preserved with silage technology as cattle ration mixture, investigate the adequacy of nutrient for the cattle after treated with addition of CPH silage, and investigate the quota of cattle treated with CPH silage addition. The research design was conducted by giving extension and field observation that was carried out at Taluditi on July to August 2015 during the activity of Student Community Service (KKN PPM UGM Unit Gorontalo 02). The secondary data was gathered from the Department of Agriculture and Plantation Pohuwatu Regency, and also supporting references. The number of respondents in each location was about 30 - 40 people. The research results showed that the silage preservation technology can be well received by the farmers. There was improvement of cattle ration nutrient supplemented with CPH compared to that of ration nutrient which was usually be used by the farmers or standard ration nutrient. The research also resulted in fresh CPH production 2,625.741 tons/year, CPH silage production 2,140.549 tons/year and load capacity + 575 heads/day weight 250-300 kg.

Isolation, characterization, and development of WRKY genes as useful genetic markers in Theobroma cacao.

PubMed

Borrone, James W; Kuhn, David N; Schnell, Raymond J

2004-08-01

There is currently an international effort in improving disease resistance and crop yield in Theobroma cacao L., an economically important crop of the tropics, using marker-assisted selection for breeding. We are developing molecular genetic markers focusing upon gene families involved with disease resistance. One such family is the WRKY proteins, which are plant-specific transcriptional factors associated with regulating defense responses to both abiotic and biotic stresses. Degenerate PCR primers were designed to the highly conserved DNA-binding domain and other conserved motifs of group I and group II, subgroups a-c, WRKY genes. Sixteen individual WRKY fragments were isolated from a mixture of T. cacao DNA using one pair of primers. Of the 16 WRKY loci investigated, seven contained single nucleotide polymorphisms within the intron as detected by sequence comparison of the PCR products. Four of these were successfully converted into molecular markers and mapped in an F2 population by capillary electrophoresis-single strand conformation polymorphism analysis. This is the first report of a pair of degenerate primers amplifying WRKY loci directly from genomic DNA and demonstrates a simple method for developing useful genetic markers from members of a large gene family. Copyright 2004 Springer-Verlag

Alpha List of Prime Contract Awards. Oct 92-Sep 93. FY93. (Sets Technology Inc. - System/Technology Development). Part 17

DTIC Science & Technology

1994-03-01

00)0oN e4 N I cacao I -4N Y 4 .0 U)nendo4)0(0 0 00a) to U)(a I)0).Ř -4 i G I ) a)CV) 0 a)Nf4r0 0) 0to n 4- 4 44-N gV24 CqIt n0I I 0)0a coo 1 Ow 0 0 0...4 0 M0 W 0 0 0 W0.-4-4W-40 -40) -4 0 M flN MN T% N lc4M-4N 1. -0 -40(0)WG CO I cacao ) 11J0 -.100) ) N N ON NON-0 (0.-. -4 - - CO) -0)ON- X- N I000...c00 W -Nu 4Wf-C) IL -4 M LLa. N C 0 0 -4 -40 o z X4 0-4-.N U c 3000 K 4N qt NY N .N az zI cacao III N 0 O 0 Ca 0 00 N0z N O I 000 UM 0I 00 11L 400 4

Mechanical stress caused by wind on leaves of Theobroma cacao: Photosynthetic, molecular, antioxidative and ultrastructural responses.

PubMed

Reis, Graciele Santos Monteiro; Almeida, Alex-Alan Furtado de; Mangabeira, Pedro Antônio Oliveira; Santos, Ivanildes Conceição Dos; Pirovani, Carlos Priminho; Ahnert, Dário

2018-01-01

Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems. Since it is a species quite intolerant to wind, it is practically impossible to implant cacao crops under full exposure to the sun, or in areas of frequent winds, without the protection of windbreaks, using arboreal species around the area of culture in the form of box. Wind can cause mechanical stimuli in plants, affecting their growth and development. The objective of this work was to evaluate the photosynthetic changes in mature leaves and the molecular, biochemical and ultrastructural changes in young and mature leaves of the CCN 51 cloned genotype of T. cacao subjected to intermittent (IW) and constant (CW) wind, with velocities of 2.5, 3.5 and 4.5 m s-1, during 3, 6 and 12 h of exposure. It was verified that CW and IW, considering different exposure times, interfered directly in stomatal conductance (gs), transpiration (E) and water use efficiency (WUE), causing a reduction of the photosynthetic rate (A) in mature leaves. In addition, the pulvinus and blade of young and mature leaves, exposed to IW and CW with different exposure times (3 and 12 h), showed marked macroscopic and microscopic mechanical injuries resulting from the constant leaf movement. At both speeds, there was rupture of the cell nuclear membrane in pulvinus and the mesophyll tissues, mainly in the young leaves. On the other hand, in young and mature leaves exposed to CW and IW at different speeds and exposure times, there was lipid peroxidation, increased activity of guaiacol (GPX) and ascorbate (APX) peroxidases in most treatments; and altered expression of transcripts of psba and psbo genes related to the phothosynthetic apparatus and Cu-Zn-sod and per genes related to antioxidative enzymes at the rate of 4.5 m s-1. Younger leaves were more intolerant to mechanical stress caused by the wind, since presented greater macro and microscopic damages and, consequently, greater molecular, biochemical and ultrastructural changes. High wind speeds can seriously compromise the development of young leaves of T. cacao plants and affect their productivity.

Genome and secretome analysis of the hemibiotrophic fungal pathogen, Moniliophthora roreri, which causes frosty pod rot disease of cacao: mechanisms of the biotrophic and necrotrophic phases.

PubMed

Meinhardt, Lyndel W; Costa, Gustavo Gilson Lacerda; Thomazella, Daniela P T; Teixeira, Paulo José P L; Carazzolle, Marcelo Falsarella; Schuster, Stephan C; Carlson, John E; Guiltinan, Mark J; Mieczkowski, Piotr; Farmer, Andrew; Ramaraj, Thiruvarangan; Crozier, Jayne; Davis, Robert E; Shao, Jonathan; Melnick, Rachel L; Pereira, Gonçalo A G; Bailey, Bryan A

2014-02-27

The basidiomycete Moniliophthora roreri is the causal agent of Frosty pod rot (FPR) disease of cacao (Theobroma cacao), the source of chocolate, and FPR is one of the most destructive diseases of this important perennial crop in the Americas. This hemibiotroph infects only cacao pods and has an extended biotrophic phase lasting up to sixty days, culminating in plant necrosis and sporulation of the fungus without the formation of a basidiocarp. We sequenced and assembled 52.3 Mb into 3,298 contigs that represent the M. roreri genome. Of the 17,920 predicted open reading frames (OFRs), 13,760 were validated by RNA-Seq. Using read count data from RNA sequencing of cacao pods at 30 and 60 days post infection, differential gene expression was estimated for the biotrophic and necrotrophic phases of this plant-pathogen interaction. The sequencing data were used to develop a genome based secretome for the infected pods. Of the 1,535 genes encoding putative secreted proteins, 1,355 were expressed in the biotrophic and necrotrophic phases. Analysis of the data revealed secretome gene expression that correlated with infection and intercellular growth in the biotrophic phase and invasive growth and plant cellular death in the necrotrophic phase. Genome sequencing and RNA-Seq was used to determine and validate the Moniliophthora roreri genome and secretome. High sequence identity between Moniliophthora roreri genes and Moniliophthora perniciosa genes supports the taxonomic relationship with Moniliophthora perniciosa and the relatedness of this fungus to other basidiomycetes. Analysis of RNA-Seq data from infected plant tissues revealed differentially expressed genes in the biotrophic and necrotrophic phases. The secreted protein genes that were upregulated in the biotrophic phase are primarily associated with breakdown of the intercellular matrix and modification of the fungal mycelia, possibly to mask the fungus from plant defenses. Based on the transcriptome data, the upregulated secreted proteins in the necrotrophic phase are hypothesized to be actively attacking the plant cell walls and plant cellular components resulting in necrosis. These genes are being used to develop a new understanding of how this disease interaction progresses and to identify potential targets to reduce the impact of this devastating disease.

Genome and secretome analysis of the hemibiotrophic fungal pathogen, Moniliophthora roreri, which causes frosty pod rot disease of cacao: mechanisms of the biotrophic and necrotrophic phases

PubMed Central

2014-01-01

Background The basidiomycete Moniliophthora roreri is the causal agent of Frosty pod rot (FPR) disease of cacao (Theobroma cacao), the source of chocolate, and FPR is one of the most destructive diseases of this important perennial crop in the Americas. This hemibiotroph infects only cacao pods and has an extended biotrophic phase lasting up to sixty days, culminating in plant necrosis and sporulation of the fungus without the formation of a basidiocarp. Results We sequenced and assembled 52.3 Mb into 3,298 contigs that represent the M. roreri genome. Of the 17,920 predicted open reading frames (OFRs), 13,760 were validated by RNA-Seq. Using read count data from RNA sequencing of cacao pods at 30 and 60 days post infection, differential gene expression was estimated for the biotrophic and necrotrophic phases of this plant-pathogen interaction. The sequencing data were used to develop a genome based secretome for the infected pods. Of the 1,535 genes encoding putative secreted proteins, 1,355 were expressed in the biotrophic and necrotrophic phases. Analysis of the data revealed secretome gene expression that correlated with infection and intercellular growth in the biotrophic phase and invasive growth and plant cellular death in the necrotrophic phase. Conclusions Genome sequencing and RNA-Seq was used to determine and validate the Moniliophthora roreri genome and secretome. High sequence identity between Moniliophthora roreri genes and Moniliophthora perniciosa genes supports the taxonomic relationship with Moniliophthora perniciosa and the relatedness of this fungus to other basidiomycetes. Analysis of RNA-Seq data from infected plant tissues revealed differentially expressed genes in the biotrophic and necrotrophic phases. The secreted protein genes that were upregulated in the biotrophic phase are primarily associated with breakdown of the intercellular matrix and modification of the fungal mycelia, possibly to mask the fungus from plant defenses. Based on the transcriptome data, the upregulated secreted proteins in the necrotrophic phase are hypothesized to be actively attacking the plant cell walls and plant cellular components resulting in necrosis. These genes are being used to develop a new understanding of how this disease interaction progresses and to identify potential targets to reduce the impact of this devastating disease. PMID:24571091

Differential expression of the lethal gene Luteus-Pa in cacao of the Parinari series.

PubMed

Rehem, B C; Almeida, A-A F; Figueiredo, G S F; Gesteira, A S; Santos, S C; Corrêa, R X; Yamada, M M; Valle, R R

2016-02-22

The recessive lethal character Luteus-Pa is found in cacao (Theobroma cacao) genotypes of the Parinari series (Pa) and is characterized by expression of leaf chlorosis and seedling death. Several genotypes of the Pa series are bearers of the gene responsible for the expression of the Luteus-Pa character, which can be used as a tool for determining relationships between genotypes of this group. To evaluate this phenomenon, we analyzed the differential expression of genes between mutant seedlings and wild-type hybrid Pa 30 x 169 seedlings, with the aim of elucidating the possible lethal mechanisms of the homozygous recessive character Luteus-Pa. Plant material was harvested from leaves of wild and mutant seedlings at different periods to construct a subtractive library and perform quantitative analysis using real-time PCR. The 649 sequences obtained from the subtractive library had an average length of 500 bp, forming 409 contigs. The probable proteins encoded were grouped into 10 functional categories. Data from ESTs identified genes associated with Rubisco, peroxidases, and other proteins and enzymes related to carbon assimilation, respiration, and photosystem 2. Mutant seedlings were characterized by synthesizing defective PsbO and PsbA proteins, which were overexpressed from 15 to 20 days after seedling emergence.

Characterization of the legumains encoded by the genome of Theobroma cacao L.

PubMed

Santana, Juliano Oliveira; Freire, Laís; de Sousa, Aurizangela Oliveira; Fontes Soares, Virgínia Lúcia; Gramacho, Karina Peres; Pirovani, Carlos Priminho

2016-01-01

Legumains are cysteine proteases related to plant development, protein degradation, programmed cell death, and defense against pathogens. In this study, we have identified and characterized three legumains encoded by Theobroma cacao genome through in silico analyses, three-dimensional modeling, genetic expression pattern in different tissues and as a response to the inoculation of Moniliophthora perniciosa fungus. The three proteins were named TcLEG3, TcLEG6, and TcLEG9. Histidine and cysteine residue which are part of the catalytic site were conserved among the proteins, and they remained parallel in the loop region in the 3D modeling. Three-dimensional modeling showed that the propeptide, which is located in the terminal C region of legumains blocks the catalytic cleft. Comparing dendrogram data with the relative expression analysis, indicated that TcLEG3 is related to the seed legumain group, TcLEG6 is related with the group of embryogenesis activities, and protein TcLEG9, with processes regarding the vegetative group. Furthermore, the expression analyses proposes a significant role for the three legumains during the development of Theobroma cacao and in its interaction with M. perniciosa. Copyright © 2015 Universidade Estadual de Santa Cruz, CNPJ: 40738999/0001-95. Published by Elsevier Masson SAS.. All rights reserved.

Cacao seeds are a "Super Fruit": A comparative analysis of various fruit powders and products

PubMed Central

2011-01-01

Background Numerous popular media sources have developed lists of "Super Foods" and, more recently, "Super Fruits". Such distinctions often are based on the antioxidant capacity and content of naturally occurring compounds such as polyphenols within those whole fruits or juices of the fruit which may be linked to potential health benefits. Cocoa powder and chocolate are made from an extract of the seeds of the fruit of the Theobroma cacao tree. In this study, we compared cocoa powder and cocoa products to powders and juices derived from fruits commonly considered "Super Fruits". Results Various fruit powders and retail fruit products were obtained and analyzed for antioxidant capacity (ORAC (μM TE/g)), total polyphenol content (TP (mg/g)), and total flavanol content (TF (mg/g)). Among the various powders that were tested, cocoa powder was the most concentrated source of ORAC and TF. Similarly, dark chocolate was a significantly more concentrated source of ORAC and TF than the fruit juices. Conclusions Cocoa powder and dark chocolate had equivalent or significantly greater ORAC, TP, and TF values compared to the other fruit powders and juices tested, respectively. Cacao seeds thus provide nutritive value beyond that derived from their macronutrient composition and appear to meet the popular media's definition of a "Super Fruit". PMID:21299842

Cacao seeds are a "Super Fruit": A comparative analysis of various fruit powders and products.

PubMed

Crozier, Stephen J; Preston, Amy G; Hurst, Jeffrey W; Payne, Mark J; Mann, Julie; Hainly, Larry; Miller, Debra L

2011-02-07

Numerous popular media sources have developed lists of "Super Foods" and, more recently, "Super Fruits". Such distinctions often are based on the antioxidant capacity and content of naturally occurring compounds such as polyphenols within those whole fruits or juices of the fruit which may be linked to potential health benefits. Cocoa powder and chocolate are made from an extract of the seeds of the fruit of the Theobroma cacao tree. In this study, we compared cocoa powder and cocoa products to powders and juices derived from fruits commonly considered "Super Fruits". Various fruit powders and retail fruit products were obtained and analyzed for antioxidant capacity (ORAC (μM TE/g)), total polyphenol content (TP (mg/g)), and total flavanol content (TF (mg/g)). Among the various powders that were tested, cocoa powder was the most concentrated source of ORAC and TF. Similarly, dark chocolate was a significantly more concentrated source of ORAC and TF than the fruit juices. Cocoa powder and dark chocolate had equivalent or significantly greater ORAC, TP, and TF values compared to the other fruit powders and juices tested, respectively. Cacao seeds thus provide nutritive value beyond that derived from their macronutrient composition and appear to meet the popular media's definition of a "Super Fruit".

Evaluation of soil amendments as a remediation alternative for cadmium-contaminated soils under cacao plantations.

PubMed

Chavez, E; He, Z L; Stoffella, P J; Mylavarapu, R; Li, Y; Baligar, V C

2016-09-01

Elevated plant-available cadmium (Cd) in soils results in contamination to cacao (Theobroma cacao L) beans. Effectiveness of vermicompost and zeolite in reducing available Cd in three cacao-growing soils was studied under laboratory conditions. Sorption-desorption experiments were conducted in soils and amendments. Cadmium was added at 0 or 5 mg kg(-1) (spiked), then, amendments were incorporated at 0, 0.5, or 2 %. Amended soils were incubated at room temperature for 28 days. Plant-available Cd was determined using 0.01 M CaCl2 (WSE) and Mehlich 3 (M3) extraction procedures in subsamples taken from individual bags at six time intervals. Soils and amendments displayed different sorption characteristics and a better fit was attained with Freundlich model (R (2) > 0.82). Amendments were ineffective in reducing extractable Cd in non-spiked soils. In Cd-spiked soils, vermicompost at 2 % significantly reduced WSE-Cd (P < 0.01) from 3.36, 0.54, and 0.38 mg kg(-1) to values lower that instrument's detection in all the three soils and significantly diminished M3-extractable Cd (P < 0.05) from 4.62 to 4.11 mg kg(-1) in only one soil. Vermicompost at 0.5 % significantly decreased WSE-Cd (P < 0.01) from 3.04 and 0.31 to 1.69 and 0.20 mg kg(-1), respectively, in two soils with low sorption capacity for Cd. In contrast, zeolite failed to reduce WSE- or M3-extractable Cd in all studied soils. A negative correlation occurred between soil pH and WSE-Cd (r > -0.89, P < 0.01). The decrease in WSE-Cd appears to be associated with the increase in pH of the vermicompost-amended soils.

Diversity of cacao trees in Waslala, Nicaragua: associations between genotype spectra, product quality and yield potential.

PubMed

Trognitz, Bodo; Cros, Emile; Assemat, Sophie; Davrieux, Fabrice; Forestier-Chiron, Nelly; Ayestas, Eusebio; Kuant, Aldo; Scheldeman, Xavier; Hermann, Michael

2013-01-01

The sensory quality and the contents of quality-determining chemical compounds in unfermented and fermented cocoa from 100 cacao trees (individual genotypes) representing groups of nine genotype spectra (GG), grown at smallholder plantings in the municipality of Waslala, Nicaragua, were evaluated for two successive harvest periods. Cocoa samples were fermented using a technique mimicking recommended on-farm practices. The sensory cocoa quality was assessed by experienced tasters, and seven major chemical taste compounds were quantified by near infrared spectrometry (NIRS). The association of the nine, partially admixed, genotype spectra with the analytical and sensory quality parameters was tested. The individual parameters were analyzed as a function of the factors GG and harvest (including the date of fermentation), individual trees within a single GG were used as replications. In fermented cocoa, significant GG-specific differences were observed for methylxanthines, theobromine-to-caffeine (T/C) ratio, total fat, procyanidin B5 and epicatechin, as well as the sensory attributes global score, astringency, and dry fruit aroma, but differences related to harvest were also apparent. The potential cocoa yield was also highly determined by the individual GG, although there was significant tree-to-tree variation within every single GG. Non-fermented samples showed large harvest-to-harvest variation of their chemical composition, while differences between GG were insignificant. These results suggest that selection by the genetic background, represented here by groups of partially admixed genotype spectra, would be a useful strategy toward enhancing quality and yield of cocoa in Nicaragua. Selection by the GG within the local, genetically segregating populations of seed-propagated cacao, followed by clonal propagation of best-performing individuals of the selected GG could be a viable alternative to traditional propagation of cacao by seed from open pollination. Fast and gentle air-drying of the fermented beans and their permanent dry storage were an efficient and comparatively easy precondition for high cocoa quality.

Diversity of Cacao Trees in Waslala, Nicaragua: Associations between Genotype Spectra, Product Quality and Yield Potential

PubMed Central

Trognitz, Bodo; Cros, Emile; Assemat, Sophie; Davrieux, Fabrice; Forestier-Chiron, Nelly; Ayestas, Eusebio; Kuant, Aldo; Scheldeman, Xavier; Hermann, Michael

2013-01-01

The sensory quality and the contents of quality-determining chemical compounds in unfermented and fermented cocoa from 100 cacao trees (individual genotypes) representing groups of nine genotype spectra (GG), grown at smallholder plantings in the municipality of Waslala, Nicaragua, were evaluated for two successive harvest periods. Cocoa samples were fermented using a technique mimicking recommended on-farm practices. The sensory cocoa quality was assessed by experienced tasters, and seven major chemical taste compounds were quantified by near infrared spectrometry (NIRS). The association of the nine, partially admixed, genotype spectra with the analytical and sensory quality parameters was tested. The individual parameters were analyzed as a function of the factors GG and harvest (including the date of fermentation), individual trees within a single GG were used as replications. In fermented cocoa, significant GG-specific differences were observed for methylxanthines, theobromine-to-caffeine (T/C) ratio, total fat, procyanidin B5 and epicatechin, as well as the sensory attributes global score, astringency, and dry fruit aroma, but differences related to harvest were also apparent. The potential cocoa yield was also highly determined by the individual GG, although there was significant tree-to-tree variation within every single GG. Non-fermented samples showed large harvest-to-harvest variation of their chemical composition, while differences between GG were insignificant. These results suggest that selection by the genetic background, represented here by groups of partially admixed genotype spectra, would be a useful strategy toward enhancing quality and yield of cocoa in Nicaragua. Selection by the GG within the local, genetically segregating populations of seed-propagated cacao, followed by clonal propagation of best-performing individuals of the selected GG could be a viable alternative to traditional propagation of cacao by seed from open pollination. Fast and gentle air-drying of the fermented beans and their permanent dry storage were an efficient and comparatively easy precondition for high cocoa quality. PMID:23349790

Spatial and temporal effects of drought on soil CO2 efflux in a cacao agroforestry system in Sulawesi, Indonesia

NASA Astrophysics Data System (ADS)

van Straaten, O.; Veldkamp, E.; Köhler, M.; Anas, I.

2010-04-01

Climate change induced droughts pose a serious threat to ecosystems across the tropics and sub-tropics, particularly to those areas not adapted to natural dry periods. In order to study the vulnerability of cacao (Theobroma cacao) - Gliricidia sepium agroforestry plantations to droughts a large scale throughfall displacement roof was built in Central Sulawesi, Indonesia. In this 19-month experiment, we compared soil surface CO2 efflux (soil respiration) from three roof plots with three adjacent control plots. Soil respiration rates peaked at intermediate soil moisture conditions and decreased under increasingly dry conditions (drought induced), or increasingly wet conditions (as evidenced in control plots). The roof plots exhibited a slight decrease in soil respiration compared to the control plots (average 13% decrease). The strength of the drought effect was spatially variable - while some measurement chamber sites reacted strongly (responsive) to the decrease in soil water content (up to R2=0.70) (n=11), others did not react at all (non-responsive) (n=7). A significant correlation was measured between responsive soil respiration chamber sites and sap flux density ratios of cacao (R=0.61) and Gliricidia (R=0.65). Leaf litter CO2 respiration decreased as conditions became drier. The litter layer contributed approximately 3-4% of the total CO2 efflux during dry periods and up to 40% during wet periods. Within days of roof opening soil CO2 efflux rose to control plot levels. Thereafter, CO2 efflux remained comparable between roof and control plots. The cumulative effect on soil CO2 emissions over the duration of the experiment was not significantly different: the control plots respired 11.1±0.5 Mg C ha-1 yr-1, while roof plots respired 10.5±0.5 Mg C ha-1 yr-1. The relatively mild decrease measured in soil CO2 efflux indicates that this agroforestry ecosystem is capable of mitigating droughts with only minor stress symptoms.

Antimicrobial properties of two novel peptides derived from Theobroma cacao osmotin.

PubMed

Falcao, Loeni L; Silva-Werneck, Joseilde O; Ramos, Alessandra de R; Martins, Natalia F; Bresso, Emmanuel; Rodrigues, Magali A; Bemquerer, Marcelo P; Marcellino, Lucilia H

2016-05-01

The osmotin proteins of several plants display antifungal activity, which can play an important role in plant defense against diseases. Thus, this protein can be useful as a source for biotechnological strategies aiming to combat fungal diseases. In this work, we analyzed the antifungal activity of a cacao osmotin-like protein (TcOsm1) and of two osmotin-derived synthetic peptides with antimicrobial features, differing by five amino acids residues at the N-terminus. Antimicrobial tests showed that TcOsm1 expressed in Escherichia coli inhibits the growth of Moniliophthora perniciosa mycelium and Pichia pastoris X-33 in vitro. The TcOsm1-derived peptides, named Osm-pepA (H-RRLDRGGVWNLNVNPGTTGARVWARTK-NH2), located at R23-K49, and Osm-pepB (H-GGVWNLNVNPGTTGARVWARTK-NH2), located at G28-K49, inhibited growth of yeasts (Saccharomyces cerevisiae S288C and Pichia pastoris X-33) and spore germination of the phytopathogenic fungi Fusarium f. sp. glycines and Colletotrichum gossypi. Osm-pepA was more efficient than Osm-pepB for S. cerevisiae (MIC=40μM and MIC=127μM, respectively), as well as for P. pastoris (MIC=20μM and MIC=127μM, respectively). Furthermore, the peptides presented a biphasic performance, promoting S. cerevisiae growth in doses around 5μM and inhibiting it at higher doses. The structural model for these peptides showed that the five amino acids residues, RRLDR at Osm-pepA N-terminus, significantly affect the tertiary structure, indicating that this structure is important for the peptide antimicrobial potency. This is the first report of development of antimicrobial peptides from T. cacao. Taken together, the results indicate that the cacao osmotin and its derived peptides, herein studied, are good candidates for developing biotechnological tools aiming to control phytopathogenic fungi. Copyright © 2016 Elsevier Inc. All rights reserved.

Bacillus subtilis and Enterobacter cloacae endophytes from healthy Theobroma cacao L. trees can systemically colonize seedlings and promote growth.

PubMed

Leite, Hianna Almeida Câmara; Silva, Anderson Barbosa; Gomes, Fábio Pinto; Gramacho, Karina Peres; Faria, José Cláudio; de Souza, Jorge Teodoro; Loguercio, Leandro Lopes

2013-03-01

Clonal genotypes resistant to fungal diseases are an important component of the cocoa production system in southeastern Bahia state (Brazil), so that technologies for faster production of stronger and healthier plantlets are highly desirable. In this study, the effects of inoculated bacterial endophytes isolated from healthy adult cacao plants on seedlings, and aspects related to inoculation methods, colonization patterns, and photosynthesis were investigated. Sequencing of 16S rRNA, hsp-60, and rpo-B genes placed the wild-type isolates within the species Enterobacter cloacae (isolates 341 and 344) and Bacillus subtilis (isolate 629). Spontaneous rifampicin-resistant (rif(R)) variants for 344 were also produced and tested. Endophytic application was either by immersion of surface sterilized seeds in bacterial suspensions or direct inoculation into soil, 20 days after planting non-inoculated seeds into pots. Results from in vitro recovery of inoculated isolates showed that the wild-type endophytes and rif(R) variants systemically colonized the entire cacao seedlings in 15-20 days, regardless of the inoculation method. Some endophytic treatments showed significant increases in seedlings' height, number of leaves, and dry matter. Inoculation methods affected the combined application of endophytes, which maintained the growth-promotion effects, but not in the same manner as in single applications. Interestingly, the 344-3.2 rif(R) variant showed improved performance in relation to both the wild type and another related variant. Photosynthetic rates and stomatal conductance increased significantly for some endophytic treatments, being partially associated with effects on growth and affected by the inoculation method. The results suggest that E. cloacae and B. subtilis endophytes from healthy adult plants (not transmitted by seeds) were able to promote vegetative growth on cacao seedlings. The development of products for large-scale use in seedlings/plantlets production systems was discussed.

[Effect of the harvest season on the composition of raw and fermented cotyledons of 2 varieties of cacao and shell fractions].

PubMed

de Dios Alvarado, J; Villacís, F E; Zamora, G F

1983-06-01

A study was carried out wherein during the period August 1979 to January 1980, samples of raw and fermented cacao were analyzed monthly. These included two varieties: Arriba, taken from a farm in "Quevedo", and the EET-19, grown in "Pichilingüe" by the Instituto Nacional de Investigaciones Agropecuarias (INIAP). Taking the ear of cacao as a basis, the weight of its main parts was determined. The proximal composition was established in the cotyledons, with significant statistical differences in regard to moisture, protein, and ether extract content according to the month of harvest. As to the fermentation process, differences in moisture, ether extract and ash content were detected; differences in the ether extract and ash content were found between the two varieties. The fat extracted from the cotyledons presented different iodine, saponification and acidity index values between the raw and fermented samples, but none were determined between the varieties; as far as the month of harvest is concerned, differences in the acidity index were observed. The percentage composition of the main fatty acids is reported (palmitic, stearic, oleic, and linoleic acids). In order to suggest possible industrial ways of utilizing the cacao shell by-product which is discarded by the shelling machine, the chemical characteristics of five fractions were determined based on the functioning of the shelling machine. The moisture, protein, ether extract, ash, crude fiber, theobromine, and caffeine contents varied among the fractions, and it was dependent on the broken "nibs" content. Differences in the protein, ether extract, and ash content, according to the months of production, were found. Obviously, the high fat content in fractions A (fine dust) and B (fine ground), which varied from 30 to 11 g/100 g, merits its extraction; the remainder meal has a valuable protein and alkaloid content. The chemical characteristics of the fat extracted from the shell of two fractions were similar to the fat extracted from the cotyledons.

Green synthesis of Pd/CuO nanoparticles by Theobroma cacao L. seeds extract and their catalytic performance for the reduction of 4-nitrophenol and phosphine-free Heck coupling reaction under aerobic conditions.

PubMed

Nasrollahzadeh, Mahmoud; Sajadi, S Mohammad; Rostami-Vartooni, Akbar; Bagherzadeh, Mojtaba

2015-06-15

We report the green synthesis of palladium/CuO nanoparticles (Pd/CuO NPs) using Theobroma cacao L. seeds extract and their catalytic activity for the reduction of 4-nitrophenol and Heck coupling reaction under aerobic conditions. The catalyst was characterized using the powder XRD, TEM, EDS, UV-vis and FT-IR. This method has the advantages of high yields, elimination of surfactant, ligand and homogeneous catalysts, simple methodology and easy work up. The catalyst can be recovered from the reaction mixture and reused several times without any significant loss of catalytic activity. Copyright © 2015 Elsevier Inc. All rights reserved.

Crinipellis brasiliensis, a new species based on morphological and molecular data.

PubMed

de Arruda, Maricília C C; Sepulveda, German F; Miller, Robert N G; Ferreira, Marisa A S V; Santiago, Denise V R; Resende, Mário Lúcio V; Dianese, José Carmine; Felipe, Maria Sueli S

2005-01-01

Crinipellis perniciosa infects a diversity of hosts causing severe damage to T. cacao production in many Brazilian growing regions. We compared isolates of Crinipellis from different geographic origins and hosts in Brazil by structural analysis using light (LM) and scanning electronic microscopy (SEM), as well as RFLP and sequence data based on the nuclear rDNA ITS region. Statistical analyses of morphometric data of basidia and basidiospores revealed a distinct group of isolates of Crinipellis obtained from Heteropterys acutifolia when compared to representatives from Theobroma cacao, Solanum lycocarpum and Heteropterys nervosa. A similar distinction also was observed based on sequence data of the ITS region such that combined results allowed for the segregation of a new species within the genus Crinipellis.

High-Resolution Transcript Profiling of the Atypical Biotrophic Interaction between Theobroma cacao and the Fungal Pathogen Moniliophthora perniciosa[C][W][OPEN

PubMed Central

Teixeira, Paulo José Pereira Lima; Thomazella, Daniela Paula de Toledo; Reis, Osvaldo; do Prado, Paula Favoretti Vital; do Rio, Maria Carolina Scatolin; Fiorin, Gabriel Lorencini; José, Juliana; Costa, Gustavo Gilson Lacerda; Negri, Victor Augusti; Mondego, Jorge Maurício Costa; Mieczkowski, Piotr; Pereira, Gonçalo Amarante Guimarães

2014-01-01

Witches’ broom disease (WBD), caused by the hemibiotrophic fungus Moniliophthora perniciosa, is one of the most devastating diseases of Theobroma cacao, the chocolate tree. In contrast to other hemibiotrophic interactions, the WBD biotrophic stage lasts for months and is responsible for the most distinctive symptoms of the disease, which comprise drastic morphological changes in the infected shoots. Here, we used the dual RNA-seq approach to simultaneously assess the transcriptomes of cacao and M. perniciosa during their peculiar biotrophic interaction. Infection with M. perniciosa triggers massive metabolic reprogramming in the diseased tissues. Although apparently vigorous, the infected shoots are energetically expensive structures characterized by the induction of ineffective defense responses and by a clear carbon deprivation signature. Remarkably, the infection culminates in the establishment of a senescence process in the host, which signals the end of the WBD biotrophic stage. We analyzed the pathogen’s transcriptome in unprecedented detail and thereby characterized the fungal nutritional and infection strategies during WBD and identified putative virulence effectors. Interestingly, M. perniciosa biotrophic mycelia develop as long-term parasites that orchestrate changes in plant metabolism to increase the availability of soluble nutrients before plant death. Collectively, our results provide unique insight into an intriguing tropical disease and advance our understanding of the development of (hemi)biotrophic plant-pathogen interactions. PMID:25371547

Comparative evaluation of total RNA extraction methods in Theobroma cacao using shoot apical meristems.

PubMed

Silva, D V; Branco, S M J; Holanda, I S A; Royaert, S; Motamayor, J C; Marelli, J P; Corrêa, R X

2016-03-04

Theobroma cacao is a species of great economic importance with its beans used for chocolate production. The tree has been a target of various molecular studies. It contains many polyphenols, which complicate the extraction of nucleic acids with the extraction protocols requiring a large amount of plant material. These issues, therefore, necessitate the optimization of the protocols. The aim of the present study was to evaluate different methods for extraction of total RNA from shoot apical meristems of T. cacao 'CCN 51' and to assess the influence of storage conditions for the meristems on the extraction. The study also aimed to identify the most efficient protocol for RNA extraction using a small amount of plant material. Four different protocols were evaluated for RNA extraction using one shoot apical meristem per sample. Among these protocols, one that was more efficient was then tested to extract RNA using four different numbers of shoot apical meristems, subjected to three different storage conditions. The best protocol was tested for cDNA amplification using reverse transcription-polymerase chain reaction; the cDNA quality was determined to be satisfactory for molecular analyses. The study revealed that with the best RNA extraction protocol, one shoot apical meristem was sufficient for extraction of high-quality total RNA. The results obtained might enable advances in genetic analyses and molecular studies using reduced amount of plant material.

Antimicrobial activity of fermented Theobroma cacao pod husk extract.

PubMed

Santos, R X; Oliveira, D A; Sodré, G A; Gosmann, G; Brendel, M; Pungartnik, C

2014-09-26

Theobroma cacao L. contains more than 500 different chemical compounds some of which have been traditionally used for their antioxidant, anti-carcinogenic, immunomodulatory, vasodilatory, analgesic, and antimicrobial activities. Spontaneous aerobic fermentation of cacao husks yields a crude husk extract (CHE) with antimicrobial activity. CHE was fractioned by solvent partition with polar solvent extraction or by silica gel chromatography and a total of 12 sub-fractions were analyzed for chemical composition and bioactivity. CHE was effective against the yeast Saccharomyces cerevisiae and the basidiomycete Moniliophthora perniciosa. Antibacterial activity was determined using 6 strains: Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis (Gram-positive) and Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmonella choleraesuis (Gram-negative). At doses up to 10 mg/mL, CHE was not effective against the Gram-positive bacteria tested but against medically important P. aeruginosa and S. choleraesuis with a minimum inhibitory concentration (MIC) of 5.0 mg/mL. Sub-fractions varied widely in activity and strongest antibacterial activity was seen with CHE8 against S. choleraesuis (MIC of 1.0 mg/mL) and CHE9 against S. epidermidis (MIC of 2.5 mg/mL). All bioactive CHE fractions contained phenols, steroids, or terpenes, but no saponins. Fraction CHE9 contained flavonoids, phenolics, steroids, and terpenes, amino acids, and alkaloids, while CHE12 had the same compounds but lacked flavonoids.

High-resolution transcript profiling of the atypical biotrophic interaction between Theobroma cacao and the fungal pathogen Moniliophthora perniciosa.

PubMed

Teixeira, Paulo José Pereira Lima; Thomazella, Daniela Paula de Toledo; Reis, Osvaldo; do Prado, Paula Favoretti Vital; do Rio, Maria Carolina Scatolin; Fiorin, Gabriel Lorencini; José, Juliana; Costa, Gustavo Gilson Lacerda; Negri, Victor Augusti; Mondego, Jorge Maurício Costa; Mieczkowski, Piotr; Pereira, Gonçalo Amarante Guimarães

2014-11-01

Witches' broom disease (WBD), caused by the hemibiotrophic fungus Moniliophthora perniciosa, is one of the most devastating diseases of Theobroma cacao, the chocolate tree. In contrast to other hemibiotrophic interactions, the WBD biotrophic stage lasts for months and is responsible for the most distinctive symptoms of the disease, which comprise drastic morphological changes in the infected shoots. Here, we used the dual RNA-seq approach to simultaneously assess the transcriptomes of cacao and M. perniciosa during their peculiar biotrophic interaction. Infection with M. perniciosa triggers massive metabolic reprogramming in the diseased tissues. Although apparently vigorous, the infected shoots are energetically expensive structures characterized by the induction of ineffective defense responses and by a clear carbon deprivation signature. Remarkably, the infection culminates in the establishment of a senescence process in the host, which signals the end of the WBD biotrophic stage. We analyzed the pathogen's transcriptome in unprecedented detail and thereby characterized the fungal nutritional and infection strategies during WBD and identified putative virulence effectors. Interestingly, M. perniciosa biotrophic mycelia develop as long-term parasites that orchestrate changes in plant metabolism to increase the availability of soluble nutrients before plant death. Collectively, our results provide unique insight into an intriguing tropical disease and advance our understanding of the development of (hemi)biotrophic plant-pathogen interactions. © 2014 American Society of Plant Biologists. All rights reserved.

Oral Supplementation with Cocoa Extract Reduces UVB-Induced Wrinkles in Hairless Mouse Skin.

PubMed

Kim, Jong-Eun; Song, Dasom; Kim, Junil; Choi, Jina; Kim, Jong Rhan; Yoon, Hyun-Sun; Bae, Jung-Soo; Han, Mira; Lee, Sein; Hong, Ji Sun; Song, Dayoung; Kim, Seong-Jin; Son, Myoung-Jin; Choi, Sang-Woon; Chung, Jin Ho; Kim, Tae-Aug; Lee, Ki Won

2016-05-01

Cacao beans contain various bioactive phytochemicals that could modify the pathogeneses of certain diseases. Here, we report that oral administration of cacao powder (CP) attenuates UVB-induced skin wrinkling by the regulation of genes involved in dermal matrix production and maintenance. Transcriptome analysis revealed that 788 genes are down- or upregulated in the CP supplemented group, compared with the UVB-irradiated mouse skin controls. Among the differentially expressed genes, cathepsin G and serpin B6c play important roles in UVB-induced skin wrinkle formation. Gene regulatory network analysis also identified several candidate regulators responsible for the protective effects of CP supplementation against UVB-induced skin damage. CP also elicited antiwrinkle effects via inhibition of UVB-induced matrix metalloproteinases-1 expression in both the human skin equivalent model and human dermal fibroblasts. Inhibition of UVB-induced activator protein-1 via CP supplementation is likely to affect the expression of matrix metalloproteinases-1. CP supplementation also downregulates the expression of cathepsin G in human dermal fibroblasts. 5-(3',4'-Dihydroxyphenyl)-γ-valerolactone, a major in vivo metabolite of CP, showed effects similar to CP supplementation. These results suggest that cacao extract may offer a protective effect against photoaging by inhibiting the breakdown of dermal matrix, which leads to an overall reduction in wrinkle formation. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

21 CFR 163.130 - Milk chocolate.

Code of Federal Regulations, 2014 CFR

2014-04-01

... carbohydrate sweeteners, and may contain one or more of the other optional ingredients specified in paragraph... used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners; (3) Spices, natural and artificial...

21 CFR 163.130 - Milk chocolate.

Code of Federal Regulations, 2012 CFR

2012-04-01

... carbohydrate sweeteners, and may contain one or more of the other optional ingredients specified in paragraph... used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners; (3) Spices, natural and artificial...

21 CFR 163.130 - Milk chocolate.

Code of Federal Regulations, 2013 CFR

2013-04-01

... carbohydrate sweeteners, and may contain one or more of the other optional ingredients specified in paragraph... used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners; (3) Spices, natural and artificial...

Caffeine Confusion

MedlinePlus

... KA-feen) is a natural chemical found in tea leaves, coffee beans, cacao (the stuff used to ... many common foods and drinks, such as coffee, tea, hot cocoa, soda, chocolate, and some medicines . When ...

CACAO: A project for a laboratory for the production and characterization of thin radioactive layers

NASA Astrophysics Data System (ADS)

Bacri, C. O.; Petitbon, V.; Pierre, S.; Cacao Group

2010-02-01

CACAO, Chimie des Actinides et Cibles radioActives à Orsay (actinide chemistry and radioactive targets at Orsay), is a project under construction that consists of the installation of a hot laboratory dedicated to the production and characterization of thin radioactive layers. The project aims to be a joint CNRS-CEA national laboratory to overcome difficulties related mainly to safety issues and to the lack of knowledge and potential manpower. The first goal is to fulfill, at least, the needs of the whole French community, and to be able to coordinate the different activities related to radioactive targets. For this purpose, itis important to be complementary to already existing international installations. Inside this framework, it will of course be possible to produce and/or characterize targets for other users.

21 CFR 163.123 - Sweet chocolate.

Code of Federal Regulations, 2014 CFR

2014-04-01

... chocolate liquor with one or more optional nutritive carbohydrate sweeteners, and may contain one or more of... following safe and suitable ingredients may be used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners...

21 CFR 163.123 - Sweet chocolate.

Code of Federal Regulations, 2013 CFR

2013-04-01

... chocolate liquor with one or more optional nutritive carbohydrate sweeteners, and may contain one or more of... following safe and suitable ingredients may be used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners...

21 CFR 163.123 - Sweet chocolate.

Code of Federal Regulations, 2012 CFR

2012-04-01

... chocolate liquor with one or more optional nutritive carbohydrate sweeteners, and may contain one or more of... following safe and suitable ingredients may be used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners...

Effect of Soil pH Increase by Biochar on NO, N2O and N2 Production during Denitrification in Acid Soils

PubMed Central

Obia, Alfred; Cornelissen, Gerard; Mulder, Jan; Dörsch, Peter

2015-01-01

Biochar (BC) application to soil suppresses emission of nitrous- (N2O) and nitric oxide (NO), but the mechanisms are unclear. One of the most prominent features of BC is its alkalizing effect in soils, which may affect denitrification and its product stoichiometry directly or indirectly. We conducted laboratory experiments with anoxic slurries of acid Acrisols from Indonesia and Zambia and two contrasting BCs produced locally from rice husk and cacao shell. Dose-dependent responses of denitrification and gaseous products (NO, N2O and N2) were assessed by high-resolution gas kinetics and related to the alkalizing effect of the BCs. To delineate the pH effect from other BC effects, we removed part of the alkalinity by leaching the BCs with water and acid prior to incubation. Uncharred cacao shell and sodium hydroxide (NaOH) were also included in the study. The untreated BCs suppressed N2O and NO and increased N2 production during denitrification, irrespective of the effect on denitrification rate. The extent of N2O and NO suppression was dose-dependent and increased with the alkalizing effect of the two BC types, which was strongest for cacao shell BC. Acid leaching of BC, which decreased its alkalizing effect, reduced or eliminated the ability of BC to suppress N2O and NO net production. Just like untreated BCs, NaOH reduced net production of N2O and NO while increasing that of N2. This confirms the importance of altered soil pH for denitrification product stoichiometry. Addition of uncharred cacao shell stimulated denitrification strongly due to availability of labile carbon but only minor effects on the product stoichiometry of denitrification were found, in accordance with its modest effect on soil pH. Our study indicates that stimulation of denitrification was mainly due to increases in labile carbon whereas change in product stoichiometry was mainly due to a change in soil pH. PMID:26397367

Effect of Soil pH Increase by Biochar on NO, N2O and N2 Production during Denitrification in Acid Soils.

PubMed

Obia, Alfred; Cornelissen, Gerard; Mulder, Jan; Dörsch, Peter

2015-01-01

Biochar (BC) application to soil suppresses emission of nitrous- (N2O) and nitric oxide (NO), but the mechanisms are unclear. One of the most prominent features of BC is its alkalizing effect in soils, which may affect denitrification and its product stoichiometry directly or indirectly. We conducted laboratory experiments with anoxic slurries of acid Acrisols from Indonesia and Zambia and two contrasting BCs produced locally from rice husk and cacao shell. Dose-dependent responses of denitrification and gaseous products (NO, N2O and N2) were assessed by high-resolution gas kinetics and related to the alkalizing effect of the BCs. To delineate the pH effect from other BC effects, we removed part of the alkalinity by leaching the BCs with water and acid prior to incubation. Uncharred cacao shell and sodium hydroxide (NaOH) were also included in the study. The untreated BCs suppressed N2O and NO and increased N2 production during denitrification, irrespective of the effect on denitrification rate. The extent of N2O and NO suppression was dose-dependent and increased with the alkalizing effect of the two BC types, which was strongest for cacao shell BC. Acid leaching of BC, which decreased its alkalizing effect, reduced or eliminated the ability of BC to suppress N2O and NO net production. Just like untreated BCs, NaOH reduced net production of N2O and NO while increasing that of N2. This confirms the importance of altered soil pH for denitrification product stoichiometry. Addition of uncharred cacao shell stimulated denitrification strongly due to availability of labile carbon but only minor effects on the product stoichiometry of denitrification were found, in accordance with its modest effect on soil pH. Our study indicates that stimulation of denitrification was mainly due to increases in labile carbon whereas change in product stoichiometry was mainly due to a change in soil pH.

Effects of habitat management on different feeding guilds of herbivorous insects in cacao agroforestry systems.

PubMed

Novais, Samuel M A; Macedo-Reis, Luiz E; DaRocha, Wesley D; Neves, Frederico S

2016-06-01

Human pressure on natural habitats increases the importance of agroforests for biodiversity conservation. The objective of this study was to evaluate the role of cacao traditional cultivation system (CTCS) on the conservation of the herbivorous insect community when compared with a monodominant rubber agroforest, a type of agricultural system for cacao cultivation. The insects were sampled in three habitats in Southeastern Bahia, Brazil: native forests, CTCS and rubber agroforests. In each habitat, 18 plots of 10 m2 were established, and the structural measures were collected and herbivorous insects were sampled with a Malaise/window trap. The diversity of folivorous decreased with the simplification of vegetation structure, but species composition was similar among habitats. In addition to a decrease in the availability of resources in monodominant rubber agroforests, the latex present in these systems have limited the occurrence of species that cannot circumvent latex toxicity. The diversity of sap-sucking insects was similar among habitats, but species composition was similar only in the CTCS and native forest, and it was different in the rubber agroforest. We observed turnover and a higher frequency of individuals of the family Psyllidae in the rubber agroforest. The biology and behavior of Psyllids and absence of natural enemies enable their diversity to increase when they are adapted to a new host. We observed a shift in the composition of xylophagous insects in the rubber agroforest compared to that in other habitats. Moreover, this agroforest has low species richness, but high individual abundance. Latex extraction is likely an important additional source of volatile compounds discharged into the environment, and it increases the attraction and recruitment of coleoborers to these sites. We concluded that CTCS has an herbivorous insect community with a structure similar to the community found in native forests of the region, and they present a more interesting conservation strategy when compared to rubber agroforests. We also emphasized the potential risk of local pest outbreaks in rubber agroforests for both the rubber and associated cacao trees.

The mitochondrial genome of Moniliophthora roreri, the frosty pod rot pathogen of cacao.

PubMed

Costa, Gustavo G L; Cabrera, Odalys G; Tiburcio, Ricardo A; Medrano, Francisco J; Carazzolle, Marcelo F; Thomazella, Daniela P T; Schuster, Stephen C; Carlson, John E; Guiltinan, Mark J; Bailey, Bryan A; Mieczkowski, Piotr; Pereira, Gonçalo A G; Meinhardt, Lyndel W

2012-05-01

In this study, we report the sequence of the mitochondrial (mt) genome of the Basidiomycete fungus Moniliophthora roreri, which is the etiologic agent of frosty pod rot of cacao (Theobroma cacao L.). We also compare it to the mtDNA from the closely-related species Moniliophthora perniciosa, which causes witches' broom disease of cacao. The 94 Kb mtDNA genome of M. roreri has a circular topology and codes for the typical 14 mt genes involved in oxidative phosphorylation. It also codes for both rRNA genes, a ribosomal protein subunit, 13 intronic open reading frames (ORFs), and a full complement of 27 tRNA genes. The conserved genes of M. roreri mtDNA are completely syntenic with homologous genes of the 109 Kb mtDNA of M. perniciosa. As in M. perniciosa, M. roreri mtDNA contains a high number of hypothetical ORFs (28), a remarkable feature that make Moniliophthoras the largest reservoir of hypothetical ORFs among sequenced fungal mtDNA. Additionally, the mt genome of M. roreri has three free invertron-like linear mt plasmids, one of which is very similar to that previously described as integrated into the main M. perniciosa mtDNA molecule. Moniliophthora roreri mtDNA also has a region of suspected plasmid origin containing 15 hypothetical ORFs distributed in both strands. One of these ORFs is similar to an ORF in the mtDNA gene encoding DNA polymerase in Pleurotus ostreatus. The comparison to M. perniciosa showed that the 15 Kb difference in mtDNA sizes is mainly attributed to a lower abundance of repetitive regions in M. roreri (5.8 Kb vs 20.7 Kb). The most notable differences between M. roreri and M. perniciosa mtDNA are attributed to repeats and regions of plasmid origin. These elements might have contributed to the rapid evolution of mtDNA. Since M. roreri is the second species of the genus Moniliophthora whose mtDNA genome has been sequenced, the data presented here contribute valuable information for understanding the evolution of fungal mt genomes among closely-related species. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

Comparative Analysis of Expressed Genes from Cacao Meristems Infected by Moniliophthora perniciosa

PubMed Central

Gesteira, Abelmon S.; Micheli, Fabienne; Carels, Nicolas; Da Silva, Aline C.; Gramacho, Karina P.; Schuster, Ivan; Macêdo, Joci N.; Pereira, Gonçalo A. G.; Cascardo, Júlio C. M.

2007-01-01

Background and Aims Witches' broom disease is caused by the hemibiotrophic basidiomycete Moniliophthora perniciosa, and is one of the most important diseases of cacao in the western hemisphere. Because very little is known about the global process of such disease development, expressed sequence tags (ESTs) were used to identify genes expressed during the Theobroma cacao–Moniliophthora perniciosa interaction. Methods Two cDNA libraries corresponding to the resistant (RT) and susceptible (SP) cacao–M. perniciosa interactions were constructed from total RNA, using the DB SMART Creator cDNA library kit (Clontech). Clones were randomly selected, sequenced from the 5′ end and analysed using bioinformatics tools including in silico analysis of the differential gene expression. Key Results A total of 6884 ESTs were generated from the RT and SP cDNA libraries. These ESTs were composed of 2585 singlets and 341 contigs for a total of 2926 non-redundant sequences. The redundancy of the libraries was low and their specificity high when compared with the few other cacao libraries already published. Sequence analysis allowed the assignment of a putative functional category for 54 % of sequences, whereas approx. 22 % of sequences corresponded to unknown function and approx. 24 % of sequences did not show any significant similarity with other proteins present in the database. Despite the similar overall distribution of the sequences in functional categories between the two libraries, qualitative differences were observed. Genes involved during the defence response to pathogen infection or in programmed cell death were identified, such as pathogenesis related-proteins, trypsin inhibitor or oxalate oxidase, and some of them showed an in silico differential expression between the resistant and the susceptible interactions. Conclusions As far as is known this is the first EST resource from the cacao–M. perniciosa interaction and it is believed that it will provide a significant contribution to the understanding of the molecular mechanisms of the resistance and susceptibility of cacao to M. perniciosa, to develop strategies to control witches broom, and as a source of polymorphism for molecular marker development and marker-assisted selection. PMID:17557832

40 CFR 180.145 - Fluorine compounds; tolerances for residues.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., postharvest 15.0 Barley, pearled barley, postharvest 45.0 Cattle, meat, dried 40 Cheese 5.0 Cacao bean, roasted bean, postharvest 20 Coconut, postharvest 40 Coffee, bean, green, postharvest 15 Corn, field...

40 CFR 180.145 - Fluorine compounds; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., postharvest 15.0 Barley, pearled barley, postharvest 45.0 Cattle, meat, dried 40 Cheese 5.0 Cacao bean, roasted bean, postharvest 20 Coconut, postharvest 40 Coffee, bean, green, postharvest 15 Corn, field...

40 CFR 180.145 - Fluorine compounds; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., postharvest 15.0 Barley, pearled barley, postharvest 45.0 Cattle, meat, dried 40 Cheese 5.0 Cacao bean, roasted bean, postharvest 20 Coconut, postharvest 40 Coffee, bean, green, postharvest 15 Corn, field...

40 CFR 180.145 - Fluorine compounds; tolerances for residues.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., postharvest 15.0 Barley, pearled barley, postharvest 45.0 Cattle, meat, dried 40 Cheese 5.0 Cacao bean, roasted bean, postharvest 20 Coconut, postharvest 40 Coffee, bean, green, postharvest 15 Corn, field...

40 CFR 180.145 - Fluorine compounds; tolerances for residues.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., postharvest 15.0 Barley, pearled barley, postharvest 45.0 Cattle, meat, dried 40 Cheese 5.0 Cacao bean, roasted bean, postharvest 20 Coconut, postharvest 40 Coffee, bean, green, postharvest 15 Corn, field...

DNA and RNA polymerase activity in a Moniliophthora perniciosa mitochondrial plasmid and self-defense against oxidative stress.

PubMed

Andrade, B S; Villela-Dias, C; Gomes, D S; Micheli, F; Góes-Neto, A

2013-06-13

Moniliophthora perniciosa (Stahel) Aime and Phillips-Mora is a hemibiotrophic basidiomycete (Agaricales, Tricholomataceae) that causes witches' broom disease in cocoa (Theobroma cacao L.). This pathogen carries a stable integrated invertron-type linear plasmid in its mitochondrial genome that encodes viral-like DNA and RNA polymerases related to fungal senescence and longevity. After culturing the fungus and obtaining its various stages of development in triplicate, we carried out total RNA extraction and subsequent complementary DNA synthesis. To analyze DNA and RNA polymerase expression levels, we performed real-time reverse transcriptase polymerase chain reaction for various fungal phases of development. Our results showed that DNA and RNA polymerase gene expression in the primordium phase of M. perniciosa is related to a potential defense mechanism against T. cacao oxidative attack.

Heavy metal concentrations in cocoa beans (Theobroma cacao L.) originating from East Luwu, South Sulawesi, Indonesia

NASA Astrophysics Data System (ADS)

Assa, A.; Noor, A.; Yunus, M. R.; Misnawi; Djide, M. N.

2018-03-01

Concentrations of some heavy metals (Pb, Cu, Cd, As and Hg) were assessed for cocoa beans (Theobroma cacao L) originating from East Luwu, South Sulawesi, Indonesia after five-day fermentation. Consisting of PB 123, BR 25, and MCC 02 cocoa clones, the spectrophotometric analysis showed that concentrations of Pb, Cd, As and Hg in the cocoa beans over the three clones was below the detection limits of 0.100; 0.050, 0.010 and 0.005 mg/kg. For Cu, they were 19.343; 10.391, and 18.594 mg/kg respectively, but still below the maximum critical levels, established by the European Food Safety Authority (EFSA). Concentrations of those five heavy metals in the bean shells were found to be parallel to those in the cocoa beans, except for Pb.

Potential use of chicken egg shells and cacao pod husk as catalyst for biodiesel production

NASA Astrophysics Data System (ADS)

Andherson, T.; Rachmat, D.; Risanti, Doty D.

2018-04-01

Chicken eggshells and cacao pod husk are known as waste materials that can be used to produce catalyst of CaO and K2CO3, respectively. Waste cooking oil (WCO) is an alternative raw material to produce economical and cheaper biodiesel. WCO first undergoes a pre-treatment process that aims to remove free fatty acid (FFA). Biodiesel was produced by transesterification between triglyceride and alcohol with the presence of a catalyst. To obtain the catalyst, eggshell was calcined in a furnace at temperatures of 600 °C, 700 °C, 800 °C, and 900 °C for 6 hours. On the other hand, K2CO3 catalyst synthesis was carried out by carbonization cacao pod husk at temperature of 650°C and subsequently extracted using aquadest solvent. Catalysts were then characterized by XRD and FTIR. The biodiesel synthesis in this research was conducted at 65°C with molar ratio of 1: 6 WCO to methanol, reaction time for 1 and 2 hours, for K2CO3 and CaO, respectively. The optimum condition for the reaction using CaO was obtained for CaO that carbonized at 900°C. It produced 81.43% conversion of biodiesel. On the other hand, reaction using K2CO3 could reach a higher conversion of 85%. From the results it was clear that the biodiesel fuel production was within the recommended SNI 7182: 2015 standard.

Analysis of Gene Expression and Proteomic Profiles of Clonal Genotypes from Theobroma cacao Subjected to Soil Flooding

PubMed Central

Bertolde, Fabiana Z.; Almeida, Alex-Alan F.; Pirovani, Carlos P.

2014-01-01

Soil flooding causes changes in gene transcription, synthesis and degradation of proteins and cell metabolism. The main objective of this study was to understand the biological events of Theobroma cacao during soil flooding-induced stress, using the analyses of gene expression and activity of key enzymes involved in fermentation, as well as the identification of differentially expressed proteins by mass spectrometry in two contrasting genotypes for flooding tolerance (tolerant - TSA-792 and susceptible - TSH-774). Soil anoxia caused by flooding has led to changes in the expression pattern of genes associated with the biosynthesis of alcohol dehydrogenase (ADH), pyruvate decarboxylase (PDC) and lactate dehydrogenase (LDH) in leaves and roots of the two evaluated genotypes. Significant differences were observed between the enzyme activities of the two genotypes. Leaves and roots of the TSA-792 genotype showed higher ADH activity as compared to the TSH-774 genotype, whereas the activities of PDC and LDH have varied over the 96 h of soil flooding, being higher for TSA-792 genotype, at the initial stage, and TSH-774 genotype, at the final stage. Some of the identified proteins are those typical of the anaerobic metabolism-involved in glycolysis and alcoholic fermentation-and different proteins associated with photosynthesis, protein metabolism and oxidative stress. The ability to maintain glycolysis and induce fermentation was observed to play an important role in anoxia tolerance in cacao and may also serve to distinguish tolerant and susceptible genotypes in relation to this stressor. PMID:25289700

Analysis of gene expression and proteomic profiles of clonal genotypes from Theobroma cacao subjected to soil flooding.

PubMed

Bertolde, Fabiana Z; Almeida, Alex-Alan F; Pirovani, Carlos P

2014-01-01

Soil flooding causes changes in gene transcription, synthesis and degradation of proteins and cell metabolism. The main objective of this study was to understand the biological events of Theobroma cacao during soil flooding-induced stress, using the analyses of gene expression and activity of key enzymes involved in fermentation, as well as the identification of differentially expressed proteins by mass spectrometry in two contrasting genotypes for flooding tolerance (tolerant - TSA-792 and susceptible - TSH-774). Soil anoxia caused by flooding has led to changes in the expression pattern of genes associated with the biosynthesis of alcohol dehydrogenase (ADH), pyruvate decarboxylase (PDC) and lactate dehydrogenase (LDH) in leaves and roots of the two evaluated genotypes. Significant differences were observed between the enzyme activities of the two genotypes. Leaves and roots of the TSA-792 genotype showed higher ADH activity as compared to the TSH-774 genotype, whereas the activities of PDC and LDH have varied over the 96 h of soil flooding, being higher for TSA-792 genotype, at the initial stage, and TSH-774 genotype, at the final stage. Some of the identified proteins are those typical of the anaerobic metabolism-involved in glycolysis and alcoholic fermentation-and different proteins associated with photosynthesis, protein metabolism and oxidative stress. The ability to maintain glycolysis and induce fermentation was observed to play an important role in anoxia tolerance in cacao and may also serve to distinguish tolerant and susceptible genotypes in relation to this stressor.

Identification of a second family of genes in Moniliophthora perniciosa, the causal agent of witches' broom disease in cacao, encoding necrosis-inducing proteins similar to cerato-platanins.

PubMed

Zaparoli, Gustavo; Cabrera, Odalys García; Medrano, Francisco Javier; Tiburcio, Ricardo; Lacerda, Gustavo; Pereira, Gonçalo Guimarães

2009-01-01

The hemibiotrophic basidiomycete Moniliophthora perniciosa is the causal agent of witches' broom disease in cacao. This is a dimorphic species, with monokaryotic hyphae during the biotrophic phase, which is converted to dikaryotic mycelia during the saprophytic phase. The infection in pod is characterized by the formation of hypertrophic and hyperplasic tissues in the biotrophic phase, which is followed by necrosis and complete degradation of the organ. We found at least five sequences in the fungal genome encoding putative proteins similar to cerato-platanin (CP)-like proteins, a novel class of proteins initially found in the phytopathogen Ceratocystis fimbriata. One M. perniciosa CP gene (MpCP1) was expressed in vitro and proved to have necrosis-inducing ability in tobacco and cacao leaves. The protein is present in solution as dimers and is able to recover necrosis activity after heat treatment. Transcription analysis ex planta showed that MpCP1 is more expressed in biotrophic-like mycelia than saprotrophic mycelia. The necrosis profile presented is different from that caused by M. perniciosa necrosis and ethylene-inducing proteins (MpNEPs), another family of elicitors expressed by M. perniciosa. Remarkably, a mixture of MpCP1 with MpNEP2 led to a synergistic necrosis effect very similar to that found in naturally infected plants. This is the first report of a basidiomycete presenting both NEP1-like proteins (NLPs) and CPs in its genome.

Characterization of a stearoyl-acyl carrier protein desaturase gene family from chocolate tree, Theobroma cacao L.

PubMed

Zhang, Yufan; Maximova, Siela N; Guiltinan, Mark J

2015-01-01

In plants, the conversion of stearoyl-ACP to oleoyol-ACP is catalyzed by a plastid-localized soluble stearoyl-acyl carrier protein (ACP) desaturase (SAD). The activity of SAD significantly impacts the ratio of saturated and unsaturated fatty acids, and is thus a major determinant of fatty acid composition. The cacao genome contains eight putative SAD isoforms with high amino acid sequence similarities and functional domain conservation with SAD genes from other species. Sequence variation in known functional domains between different SAD family members suggested that these eight SAD isoforms might have distinct functions in plant development, a hypothesis supported by their diverse expression patterns in various cacao tissues. Notably, TcSAD1 is universally expressed across all the tissues, and its expression pattern in seeds is highly correlated with the dramatic change in fatty acid composition during seed maturation. Interestingly, TcSAD3 and TcSAD4 appear to be exclusively and highly expressed in flowers, functions of which remain unknown. To test the function of TcSAD1 in vivo, transgenic complementation of the Arabidopsis ssi2 mutant was performed, demonstrating that TcSAD1 successfully rescued all AtSSI2 related phenotypes further supporting the functional orthology between these two genes. The identification of the major SAD gene responsible for cocoa butter biosynthesis provides new strategies for screening for novel genotypes with desirable fatty acid compositions, and for use in breeding programs to help pyramid genes for quality and other traits such as disease resistance.

Proteomic analysis during of spore germination of Moniliophthora perniciosa, the causal agent of witches' broom disease in cacao.

PubMed

Mares, Joise Hander; Gramacho, Karina Peres; Santos, Everton Cruz; da Silva Santiago, André; Santana, Juliano Oliveira; de Sousa, Aurizângela Oliveira; Alvim, Fátima Cerqueira; Pirovani, Carlos Priminho

2017-08-17

Moniliophthora perniciosa is a phytopathogenic fungus responsible for witches' broom disease of cacao trees (Theobroma cacao L.). Understanding the molecular events during germination of the pathogen may enable the development of strategies for disease control in these economically important plants. In this study, we determined a comparative proteomic profile of M. perniciosa basidiospores during germination by two-dimensional SDS-PAGE and mass spectrometry. A total of 316 proteins were identified. Molecular changes during the development of the germinative tube were identified by a hierarchical clustering analysis based on the differential accumulation of proteins. Proteins associated with fungal filamentation, such as septin and kinesin, were detected only 4 h after germination (hag). A transcription factor related to biosynthesis of the secondary metabolite fumagillin, which can form hybrids with polyketides, was induced 2 hag, and polyketide synthase was observed 4 hag. The accumulation of ATP synthase, binding immunoglobulin protein (BiP), and catalase was validated by western blotting. In this study, we showed variations in protein expression during the early germination stages of fungus M. perniciosa. Proteins associated with fungal filamentation, and consequently with virulence, were detected in basidiospores 4 hag., for example, septin and kinesin. We discuss these results and propose a model of the germination of fungus M. perniciosa. This research can help elucidate the mechanisms underlying basic processes of host invasion and to develop strategies for control of the disease.

Bacilliform DNA-containing plant viruses in the tropics: commonalities within a genetically diverse group.

PubMed

Borah, Basanta K; Sharma, Shweta; Kant, Ravi; Johnson, A M Anthony; Saigopal, Divi Venkata Ramana; Dasgupta, Indranil

2013-10-01

Plant viruses, possessing a bacilliform shape and containing double-stranded DNA, are emerging as important pathogens in a number of agricultural and horticultural crops in the tropics. They have been reported from a large number of countries in African and Asian continents, as well as from islands from the Pacific region. The viruses, belonging to two genera, Badnavirus and Tungrovirus, within the family Caulimoviridae, have genomes displaying a common plan, yet are highly variable, sometimes even between isolates of the same virus. In this article, we summarize the current knowledge with a view to revealing the common features embedded within the genetic diversity of this group of viruses. Virus; order Unassigned; family Caulimoviridae; genera Badnavirus and Tungrovirus; species Banana streak viruses, Bougainvillea spectabilis chlorotic vein banding virus, Cacao swollen shoot virus, Citrus yellow mosaic badnavirus, Dioscorea bacilliform viruses, Rice tungro bacilliform virus, Sugarcane bacilliform viruses and Taro bacilliform virus. Bacilliform in shape; length, 60-900 nm; width, 35-50 nm; circular double-stranded DNA of approximately 7.5 kbp with one or more single-stranded discontinuities. Each virus generally limited to its own host, including banana, bougainvillea, black pepper, cacao, citrus species, Dioscorea alata, rice, sugarcane and taro. Foliar streaking in banana and sugarcane, swelling of shoots in cacao, yellow mosaic in leaves and stems in citrus, brown spot in the tubers in yam and yellow-orange discoloration and stunting in rice. http://www.dpvweb.net. 2013 BSPP and JOHN WILEY & SONS LTD

Computer Series, 114: MO Theory Made Visible.

ERIC Educational Resources Information Center

Mealli, Carlo; Proserpio, Davide M.

1990-01-01

A collection of Molecular Orbital (MO) programs that have been integrated into routines and programs to illustrate MO theory are presented. Included are discussions of Computer Aided Composition of Atomic Orbitals (CACAO) and Walsh diagrams. (CW)

Survey of commercially available chocolate- and cocoa-containing products in the United States. 2. Comparison of flavan-3-ol content with nonfat cocoa solids, total polyphenols, and percent cacao.

PubMed

Miller, Kenneth B; Hurst, W Jeffrey; Flannigan, Nancy; Ou, Boxin; Lee, C Y; Smith, Nancy; Stuart, David A

2009-10-14

A survey of a broad range of chocolate- and cocoa-containing products marketed in the United States was conducted to provide a more detailed analysis of flavan-3-ol monomers, oligomers, and polymers, which can be grouped into a class of compounds called procyanidins. Samples consisted of the three or four top-selling products within the following six categories: natural cocoa powder, unsweetened baking chocolate, dark chocolate, semisweet baking chips, milk chocolate, and chocolate syrup. Composite samples were characterized for percent fat (% fat), percent nonfat cocoa solids (% NFCS), antioxidant level by ORAC, total polyphenols, epicatechin, catechin, total monomers, and flavan-3-ol oligomers and polymers (procyanidins). On a gram weight basis epicatechin and catechin content of the products follow in decreasing order: cocoa powder > baking chocolate > dark chocolate = baking chips > milk chocolate > chocolate syrup. Analysis of the monomer and oligomer profiles within product categories shows there are two types of profiles: (1) products that have high monomers with decreasing levels of oligomers and (2) products in which the level of dimers is equal to or greater than the monomers. Results show a strong correlation (R(2) = 0.834) of epicatechin to the level of % NFCS and also very good correlations for N = 2-5 oligomers to % NFCS. A weaker correlation was observed for catechin to % NFCS (R(2) = 0.680). Other analyses show a similar high degree of correlation with epicatechin and N = 2-5 oligomers to total polyphenols, with catechin being less well correlated to total polyphenols. A lesser but still good correlation exists between the calculated percent cacao (calcd % cacao) content, a proxy for percent cacao, and these same flavanol measures, with catechin again showing a lesser degree of correlation to calcd % cacao. Principal component analysis (PCA) shows that the products group discretely into five classes: (1) cocoa powder, (2) baking chocolate, (3) dark chocolate and semisweet chips, (4) milk chocolates, and (5) syrup. PCA also shows that most factors group closely together including the antioxidant activity, total polyphenols, and the flavan-3-ol measures with the exception of catechin and % fat in the product, which group separately. Because catechin distribution appears to be different from the other flavan-3-ol measures, an analysis of the epicatechin to catechin ratio was done, indicating there is a >5-fold variation in this measure across the products studied. The cocoa-containing products tested range from cocoa powder with 227.34 +/- 17.23 mg of procyanidins per serving to 25.75 +/- 9.91 mg of procyanidins per serving for chocolate syrup. These results are discussed with respect to other studies on commercial products, the bioavailability of the flavanols, and the possible role of processing on the amount of catechin in products.

40 CFR 180.575 - Sulfuryl fluoride; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., postharvest 0.1 Barley, pearled barley, postharvest 0.05 Cacao bean, roasted bean, postharvest 0.2 Cattle, meat, dried 0.01 Cheese 2.0 Coconut, postharvest 1.0 Coffee, bean, roasted bean, postharvest 1.0 Corn...

40 CFR 180.575 - Sulfuryl fluoride; tolerances for residues.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., postharvest 0.1 Barley, pearled barley, postharvest 0.05 Cacao bean, roasted bean, postharvest 0.2 Cattle, meat, dried 0.01 Cheese 2.0 Coconut, postharvest 1.0 Coffee, bean, roasted bean, postharvest 1.0 Corn...

40 CFR 180.575 - Sulfuryl fluoride; tolerances for residues.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., postharvest 0.1 Barley, pearled barley, postharvest 0.05 Cacao bean, roasted bean, postharvest 0.2 Cattle, meat, dried 0.01 Cheese 2.0 Coconut, postharvest 1.0 Coffee, bean, roasted bean, postharvest 1.0 Corn...

40 CFR 180.575 - Sulfuryl fluoride; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., postharvest 0.1 Barley, pearled barley, postharvest 0.05 Cacao bean, roasted bean, postharvest 0.2 Cattle, meat, dried 0.01 Cheese 2.0 Coconut, postharvest 1.0 Coffee, bean, roasted bean, postharvest 1.0 Corn...

40 CFR 180.128 - Pyrethrins; tolerances for residues.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., postharvest 1.0 Apple, postharvest 1.0 Barley, grain, postharvest 3.0 Bean, succulent, postharvest 1.0..., postharvest 1.0 Buckwheat, grain, postharvest 3.0 Cacao bean, roasted bean, postharvest 1.0 Cattle, fat 1.0...

40 CFR 180.575 - Sulfuryl fluoride; tolerances for residues.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., postharvest 0.1 Barley, pearled barley, postharvest 0.05 Cacao bean, roasted bean, postharvest 0.2 Cattle, meat, dried 0.01 Cheese 2.0 Coconut, postharvest 1.0 Coffee, bean, roasted bean, postharvest 1.0 Corn...

Clinical benefit and preservation of flavonols in dark chocolate manufacturing.

PubMed

McShea, Andrew; Ramiro-Puig, Emma; Munro, Sandra B; Casadesus, Gemma; Castell, Margarida; Smith, Mark A

2008-11-01

The consumption of high-cacao-content chocolate has been associated with positive health benefits ascribed to flavanol [corrected] antioxidants derived from the ground, fermented cocoa seeds of Theobroma cacao. However, flavanols [corrected] impart a bitter, astringent flavor to foodstuffs, frequently masked in chocolates and confections by aggressive processing and adulteration with other flavors. Recent reports have implied that not all varieties of dark chocolate are created equally, and significant caveats exist regarding its potential health benefits. It is perhaps not surprising that extensive processing, dilution, and the addition of flavor modifiers may improve the palatability of chocolate, but could have negative nutritional and clinical benefits. This article examines the chemical composition of chocolate and the clinical data associated with the consumption of flavonoid-rich cocoa. We review the steps in chocolate manufacturing that directly affect the antioxidant levels in chocolate products, and the caveats associated with claims of health benefits from the consumption of dark chocolate.

Characterization of a stearoyl-acyl carrier protein desaturase gene family from chocolate tree, Theobroma cacao L

PubMed Central

Zhang, Yufan; Maximova, Siela N.; Guiltinan, Mark J.

2015-01-01

In plants, the conversion of stearoyl-ACP to oleoyol-ACP is catalyzed by a plastid-localized soluble stearoyl-acyl carrier protein (ACP) desaturase (SAD). The activity of SAD significantly impacts the ratio of saturated and unsaturated fatty acids, and is thus a major determinant of fatty acid composition. The cacao genome contains eight putative SAD isoforms with high amino acid sequence similarities and functional domain conservation with SAD genes from other species. Sequence variation in known functional domains between different SAD family members suggested that these eight SAD isoforms might have distinct functions in plant development, a hypothesis supported by their diverse expression patterns in various cacao tissues. Notably, TcSAD1 is universally expressed across all the tissues, and its expression pattern in seeds is highly correlated with the dramatic change in fatty acid composition during seed maturation. Interestingly, TcSAD3 and TcSAD4 appear to be exclusively and highly expressed in flowers, functions of which remain unknown. To test the function of TcSAD1 in vivo, transgenic complementation of the Arabidopsis ssi2 mutant was performed, demonstrating that TcSAD1 successfully rescued all AtSSI2 related phenotypes further supporting the functional orthology between these two genes. The identification of the major SAD gene responsible for cocoa butter biosynthesis provides new strategies for screening for novel genotypes with desirable fatty acid compositions, and for use in breeding programs to help pyramid genes for quality and other traits such as disease resistance. PMID:25926841

Genetic identification of Theobroma cacao L. trees with high Criollo ancestry in Soconusco, Chiapas, Mexico.

PubMed

Vázquez-Ovando, J A; Molina-Freaner, F; Nuñez-Farfán, J; Ovando-Medina, I; Salvador-Figueroa, M

2014-12-12

Criollo-type cacao trees are an important pool of genes with potential to be used in cacao breeding and selection programs. For that reason, we assessed the diversity and population structure of Criollo-type trees (108 cultivars with Criollo phenotypic characteristics and 10 Criollo references) using 12 simple sequence repeat (SSR) markers. Cultivars were selected from 7 demes in the Soconusco region of southern Mexico. SSRs amplified 74 alleles with an average of 3.6 alleles per population. The overall populations showed an average observed heterozygosity of 0.28, indicating heterozygote deficiency (average fixation index F = 0.50). However, moderate allelic diversity was found within populations (Shannon index for all populations I = 0.97). Bayesian method analysis determined 2 genetic clusters (K = 2) within individuals. In concordance, an assignment test grouped 37 multilocus genotypes (including 10 references) into a first cluster (Criollo), 54 into a second (presumably Amelonado), and 27 admixed individuals unassigned at the 90% threshold likely corresponding to the Trinitario genotype. This classification was supported by the principal coordinate analysis and analysis of molecular variance, which showed 12% of variation among populations (FST = 0.123, P < 0.0001). Sampled demes sites (1- 7) in the Soconusco region did not show any evidence of clustering by geographic location, and this was supported by the Mantel test (Rxy = 0.54, P = 0.120). Individuals with high Criollo lineage planted in Soconusco farms could be an important reservoir of genes for future breeding programs searching for fine, taste, flavor, and aroma cocoa.

Identification and analysis of the TIFY gene family in Gossypium raimondii.

PubMed

He, D H; Lei, Z P; Tang, B S; Xing, H Y; Zhao, J X; Jing, Y L

2015-08-21

The highly conserved TIFY domain is included in the TIFY protein family of transcription factors, which is important in plant development. Here, 28 TIFY family genes were identified in the Gossypium raimondii genome and classified into JAZ (15 genes), ZML (8), PPD (3), and TIFY (2). The normal (TIF[F/Y]XG) motif was dominant in the TIFY family, excluding the ZML subfamily, in which TLSFXG was prevalent. TIFY family genes were unevenly distributed in the G. raimondii genome, with TIFY clusters present on chromosome 9. Phylogenetic analysis indicated abundant variations in the G. raimondii TIFY family, which were most closely related to those in Theobroma cacao among 5 species. Exon-intron organization and intron phases were homologous within each subfamily, correlating with their phylogeny. Intra-species synteny analyses indicated that genomic duplication contributed to the expansion of the TIFY family. Inter-species synteny analyses indicated that synteny regions involved in G. raimondii TIFY family genes were also present in the comparison of G. raimondii vs Arabidopsis thaliana or T. cacao, signifying that these genes had common ancestors and play the same or similar roles in biological processes. Greater synteny was present in the comparison of G. raimondii vs T. cacao than of G. raimondii vs A. thaliana. The expression patterns of TIFY family genes were characterized and most TIFY family genes were indicated to be involved in fiber development. Our study provides new data related to the evolution of TIFYs and their role as important regulators of transcription; these data can be useful for fiber development.

Hydrogen peroxide formation in cacao tissues infected by the hemibiotrophic fungus Moniliophthora perniciosa.

PubMed

Dias, Cristiano Villela; Mendes, Juliano Sales; dos Santos, Anderson Carvalho; Pirovani, Carlos Priminho; da Silva Gesteira, Abelmon; Micheli, Fabienne; Gramacho, Karina Peres; Hammerstone, John; Mazzafera, Paulo; de Mattos Cascardo, Júlio Cézar

2011-08-01

In plant-pathogen interaction, the hydrogen peroxide (H₂O₂) may play a dual role: its accumulation inhibits the growth of biotrophic pathogens, while it could help the infection/colonization process of plant by necrotrophic pathogens. One of the possible pathways of H₂O production involves oxalic acid (Oxa) degradation by apoplastic oxalate oxidase. Here, we analyzed the production of H₂O₂, the presence of calcium oxalate (CaOx) crystals and the content of Oxa and ascorbic acid (Asa)--the main precursor of Oxa in plants--in susceptible and resistant cacao (Theobroma cacao L.) infected by the hemibiotrophic fungus Moniliophthora perniciosa. We also quantified the transcript level of ascorbate peroxidase (Apx), germin-like oxalate oxidase (Glp) and dehydroascorbate reductase (Dhar) by RT-qPCR. We report that the CaOx crystal amount and the H₂O₂ levels in the two varieties present distinct temporal and genotype-dependent patterns. Susceptible variety accumulated more CaOx crystals than the resistant one, and the dissolution of these crystals occurred in the early infection steps and in the final stage of the disease in the resistant and the susceptible variety, respectively. High expression of the Glp and accumulation of Oxa were observed in the resistant variety. The content of Asa increased in the inoculated susceptible variety, but remained constant in the resistant one. The susceptible variety presented reduced Dhar expression. The role of H₂O₂ and its formation from Oxa via Apx and Glp in resistant and susceptible variety infected by M. perniciosa were discussed. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Effect of selective logging on genetic diversity and gene flow in Cariniana legalis sampled from a cacao agroforestry system.

PubMed

Leal, J B; Santos, R P; Gaiotto, F A

2014-01-28

The fragments of the Atlantic Forest of southern Bahia have a long history of intense logging and selective cutting. Some tree species, such as jequitibá rosa (Cariniana legalis), have experienced a reduction in their populations with respect to both area and density. To evaluate the possible effects of selective logging on genetic diversity, gene flow, and spatial genetic structure, 51 C. legalis individuals were sampled, representing the total remaining population from the cacao agroforestry system. A total of 120 alleles were observed from the 11 microsatellite loci analyzed. The average observed heterozygosity (0.486) was less than the expected heterozygosity (0.721), indicating a loss of genetic diversity in this population. A high fixation index (FIS = 0.325) was found, which is possibly due to a reduction in population size, resulting in increased mating among relatives. The maximum (1055 m) and minimum (0.095 m) distances traveled by pollen or seeds were inferred based on paternity tests. We found 36.84% of unique parents among all sampled seedlings. The progenitors of the remaining seedlings (63.16%) were most likely out of the sampled area. Positive and significant spatial genetic structure was identified in this population among classes 10 to 30 m away with an average coancestry coefficient between pairs of individuals of 0.12. These results suggest that the agroforestry system of cacao cultivation is contributing to maintaining levels of diversity and gene flow in the studied population, thus minimizing the effects of selective logging.

Tradeoffs between income, biodiversity, and ecosystem functioning during tropical rainforest conversion and agroforestry intensification

PubMed Central

Steffan-Dewenter, Ingolf; Kessler, Michael; Barkmann, Jan; Bos, Merijn M.; Buchori, Damayanti; Erasmi, Stefan; Faust, Heiko; Gerold, Gerhard; Glenk, Klaus; Gradstein, S. Robbert; Guhardja, Edi; Harteveld, Marieke; Hertel, Dietrich; Höhn, Patrick; Kappas, Martin; Köhler, Stefan; Leuschner, Christoph; Maertens, Miet; Marggraf, Rainer; Migge-Kleian, Sonja; Mogea, Johanis; Pitopang, Ramadhaniel; Schaefer, Matthias; Schwarze, Stefan; Sporn, Simone G.; Steingrebe, Andrea; Tjitrosoedirdjo, Sri S.; Tjitrosoemito, Soekisman; Twele, André; Weber, Robert; Woltmann, Lars; Zeller, Manfred; Tscharntke, Teja

2007-01-01

Losses of biodiversity and ecosystem functioning due to rainforest destruction and agricultural intensification are prime concerns for science and society alike. Potentially, ecosystems show nonlinear responses to land-use intensification that would open management options with limited ecological losses but satisfying economic gains. However, multidisciplinary studies to quantify ecological losses and socioeconomic tradeoffs under different management options are rare. Here, we evaluate opposing land use strategies in cacao agroforestry in Sulawesi, Indonesia, by using data on species richness of nine plant and animal taxa, six related ecosystem functions, and on socioeconomic drivers of agroforestry expansion. Expansion of cacao cultivation by 230% in the last two decades was triggered not only by economic market mechanisms, but also by rarely considered cultural factors. Transformation from near-primary forest to agroforestry had little effect on overall species richness, but reduced plant biomass and carbon storage by ≈75% and species richness of forest-using species by ≈60%. In contrast, increased land use intensity in cacao agroforestry, coupled with a reduction in shade tree cover from 80% to 40%, caused only minor quantitative changes in biodiversity and maintained high levels of ecosystem functioning while doubling farmers' net income. However, unshaded systems further increased income by ≈40%, implying that current economic incentives and cultural preferences for new intensification practices put shaded systems at risk. We conclude that low-shade agroforestry provides the best available compromise between economic forces and ecological needs. Certification schemes for shade-grown crops may provide a market-based mechanism to slow down current intensification trends. PMID:17360392

Tradeoffs between income, biodiversity, and ecosystem functioning during tropical rainforest conversion and agroforestry intensification.

PubMed

Steffan-Dewenter, Ingolf; Kessler, Michael; Barkmann, Jan; Bos, Merijn M; Buchori, Damayanti; Erasmi, Stefan; Faust, Heiko; Gerold, Gerhard; Glenk, Klaus; Gradstein, S Robbert; Guhardja, Edi; Harteveld, Marieke; Hertel, Dietrich; Höhn, Patrick; Kappas, Martin; Köhler, Stefan; Leuschner, Christoph; Maertens, Miet; Marggraf, Rainer; Migge-Kleian, Sonja; Mogea, Johanis; Pitopang, Ramadhaniel; Schaefer, Matthias; Schwarze, Stefan; Sporn, Simone G; Steingrebe, Andrea; Tjitrosoedirdjo, Sri S; Tjitrosoemito, Soekisman; Twele, André; Weber, Robert; Woltmann, Lars; Zeller, Manfred; Tscharntke, Teja

2007-03-20

Losses of biodiversity and ecosystem functioning due to rainforest destruction and agricultural intensification are prime concerns for science and society alike. Potentially, ecosystems show nonlinear responses to land-use intensification that would open management options with limited ecological losses but satisfying economic gains. However, multidisciplinary studies to quantify ecological losses and socioeconomic tradeoffs under different management options are rare. Here, we evaluate opposing land use strategies in cacao agroforestry in Sulawesi, Indonesia, by using data on species richness of nine plant and animal taxa, six related ecosystem functions, and on socioeconomic drivers of agroforestry expansion. Expansion of cacao cultivation by 230% in the last two decades was triggered not only by economic market mechanisms, but also by rarely considered cultural factors. Transformation from near-primary forest to agroforestry had little effect on overall species richness, but reduced plant biomass and carbon storage by approximately 75% and species richness of forest-using species by approximately 60%. In contrast, increased land use intensity in cacao agroforestry, coupled with a reduction in shade tree cover from 80% to 40%, caused only minor quantitative changes in biodiversity and maintained high levels of ecosystem functioning while doubling farmers' net income. However, unshaded systems further increased income by approximately 40%, implying that current economic incentives and cultural preferences for new intensification practices put shaded systems at risk. We conclude that low-shade agroforestry provides the best available compromise between economic forces and ecological needs. Certification schemes for shade-grown crops may provide a market-based mechanism to slow down current intensification trends.

Antioxidants and Autism: Teachers' Perceptions of Behavioral Changes.

PubMed

Sadek, Amy; Berk, Lee S; Mainess, Karen; Daher, Noha S

2018-06-05

BACKGROUND- Children with Autism Spectrum Disorder (ASD) demonstrate a physiological imbalance between free radicals, resultant from oxidative stress, and antioxidants. Oxidative stress is linked to the pathogenesis of this neurocognitive disorder. The aim of this pilot feasibility study was to examine the effect of consumption of high concentration antioxidant cacao on behavior of children with ASD. METHODS- This was a 4-week pre-test post-test experimental pilot study of high antioxidant cacao and children with ASD. Participants consumed 8 squares (or 16 grams) per day of the dark chocolate which had a concentration of 70% cacao and 30% organic cane sugar (total antioxidant concentration was 8,320). The two main behavioral measures were the Aberrant Behavior Checklist- 2nd Edition and the Autism Spectrum Rating Scale which were completed by the child's teacher at baseline and end of week four. RESULTS- Sixteen participants were recruited for this study. Follow up data was available on 12 participants (9 males, 3 females, mean age of 10.9 ±3.9 years). Significant improvements on the Autism Spectrum Rating Scale were noted in Social/Communication (p=0.03, η2=0.79), Unusual Behaviors (p=0.02, η2=0.70), and Self-Regulation (p=0.04, η2=0.59). No significant changes were noted on any of the Aberrant Behavior Checklist-2 subscales (p>.05). CONCLUSION- Results from this study support the potential therapeutic benefit of antioxidants in improving social communication, unusual behaviors, and self-regulation behaviors of children with ASD. Further robust randomized controlled trials are now necessary to elaborate the validity of these findings.

Utilization of Cacao Pod Husk (Theobroma cacao l.) as Activated Carbon and Catalyst in Biodiesel Production Process from Waste Cooking Oil

NASA Astrophysics Data System (ADS)

Rachmat, Devita; Johar Mawarani, Lizda; Dewi Risanti, Doty

2018-01-01

Cocoa pod husk (Theobroma cacao l.) is a waste from cocoa beans processing. In this research we employ cocoa pod husk as activated carbon to decrease the value of FFA (Free Fatty Acid) in waste cooking oil and as K2CO3 catalyst in biodiesel production process from waste cooking oil. Cocoa pod husk was crusched and grounded into powder that passed thorugh 60 mesh-screen. As activated carbon, cocoa pod husk was firstly carbonized at three variant temperatures i.e 250°C, 300°C and 350°C. The activation process was done using HCl 2M as activator. Based on the results of XRD and FTIR, the carbonization at all variant temperatures does not cause a significant changes in terms of crystallite structure and water content. The pore of activated carbon started to form in sample that was carbonized at 350°C resulting in pore diameter of 5.14644 nm. This result was supported by the fact that the ability of this activated carbon in reducing the FFA of waste cooking oil was the most pronounced one, i.e. up to 86.7% of FFA. It was found that the performance of cocoa pod husk’s activated carbon in reducing FFA is more effective than esterification using H2SO4 which can only decrease 80.8%. On the other hand, the utilization as K2CO3 catalyst was carried out by carbonization at temperature 650°C and extraction using aquadest solvent. The extraction of cocoa pod husk produced 7.067% K2CO3 catalyst. According to RD results the fraction of K2CO3 compound from the green catalysts is the same as the commercial (SAP, 99%) that is ≥ 60%. From the obtained results, the best yield percentage was obtained using K2CO3 catalyst from cacao pod husk extract, i.e. 73-85%. To cope with biodiesel conversion efficiency, a two-step process consisting pretreatment with activated carbon carbonized at 350°C and esterification with K2CO3 from cocoa pod husk catalyst was developed. This two-step process could reach a high conversion of 85%. From the results it was clear that the produced biodiesel fuel was within the recommended SNI 7182: 2015 standard.

Cacao Flavor through Genetics – Anatomy of Fine Flavor

USDA-ARS?s Scientific Manuscript database

This presentation will discuss the transcript analysis of Moniliophthora roreri infected pods, which revealed a total of 3009 differentially expressed transcripts among resistant and susceptible clones. Comparison of the tolerant and susceptible clones by KEGG (Kyoto Encyclopedia of Genes and Genome...

Purity assessment of condensed tannin fractions by nuclear magnetic resonance (NMR) spectroscopy

USDA-ARS?s Scientific Manuscript database

Unambiguous investigation of condensed tannin (CT) structure-activity relationships in biological systems requires the use of highly enriched CT fractions of defined chemical purity. Purification of CTs from Sorghum bicolor, Trifolium repens, Theobroma cacao, Lespedeza cuneata, Lotus pedunculatus, a...

Anthelmintic activity of acetone-water extracts against Haemonchus contortus eggs: interactions between tannins and other plant secondary compounds.

PubMed

Vargas-Magaña, J J; Torres-Acosta, J F J; Aguilar-Caballero, A J; Sandoval-Castro, C A; Hoste, H; Chan-Pérez, J A

2014-12-15

This study aimed at (i) describing the effects of acetone-water extracts obtained from a range of different plant materials, on the hatching process of Haemonchus contortus eggs under in vitro conditions and (ii) identifying the role of tannins and other plant secondary compounds (PSC), on these AH effects by using polyvinylpolypyrrolidone (PVPP), an inhibitor of tannins and other polyphenols. An egg hatch assay (EHA) was used to determine the AH effect. Acetone-water (70:30) extracts from different foliages (Lysiloma latisiliquum, Laguncularia racemosa, Rizophora mangle, Avicennia germinans) and plant by-products (Theobroma cacao seed husk and pulp, and percolated Coffea arabica) were obtained. Fresh H. contortus eggs were incubated in PBS with increasing concentrations of each extract (0, 600, 1200, 2400 and 3600 μg/ml PBS). A general linear model was used to determine the dose effect of each extract. A mild ovicidal activity was only recorded for T. cacao extracts (seed husk and pulp). The main anthelmintic (AH) effect for all the extracts, except for C. arabica, was to block the eclosion of larvated eggs. The use of PVPP at 3600 μg/ml PBS showed that tannins of the L. racemosa extract were responsible for blocking eclosion of larvated eggs. Extracts of L. latisiliquum, A. germinans, T. cacao seed husk and pulp also blocked eclosion of larvated eggs but the addition of PVPP indicated that tannins were not responsible for that activity. In contrast, it suggested unfavorable interactions between polyphenols and other PSC contained in those extracts, limiting the AH effect on the egg hatching process. The present results suggest that the interactions between tannins and other PSC are complex and may reduce the AH effects against H. contortus eggs. Copyright © 2014 Elsevier B.V. All rights reserved.

Conversion of lowland tropical forests to tree cash crop plantations loses up to one-half of stored soil organic carbon

PubMed Central

van Straaten, Oliver; Corre, Marife D.; Wolf, Katrin; Tchienkoua, Martin; Cuellar, Eloy; Matthews, Robin B.; Veldkamp, Edzo

2015-01-01

Tropical deforestation for the establishment of tree cash crop plantations causes significant alterations to soil organic carbon (SOC) dynamics. Despite this recognition, the current Intergovernmental Panel on Climate Change (IPCC) tier 1 method has a SOC change factor of 1 (no SOC loss) for conversion of forests to perennial tree crops, because of scarcity of SOC data. In this pantropic study, conducted in active deforestation regions of Indonesia, Cameroon, and Peru, we quantified the impact of forest conversion to oil palm (Elaeis guineensis), rubber (Hevea brasiliensis), and cacao (Theobroma cacao) agroforestry plantations on SOC stocks within 3-m depth in deeply weathered mineral soils. We also investigated the underlying biophysical controls regulating SOC stock changes. Using a space-for-time substitution approach, we compared SOC stocks from paired forests (n = 32) and adjacent plantations (n = 54). Our study showed that deforestation for tree plantations decreased SOC stocks by up to 50%. The key variable that predicted SOC changes across plantations was the amount of SOC present in the forest before conversion—the higher the initial SOC, the higher the loss. Decreases in SOC stocks were most pronounced in the topsoil, although older plantations showed considerable SOC losses below 1-m depth. Our results suggest that (i) the IPCC tier 1 method should be revised from its current SOC change factor of 1 to 0.6 ± 0.1 for oil palm and cacao agroforestry plantations and 0.8 ± 0.3 for rubber plantations in the humid tropics; and (ii) land use management policies should protect natural forests on carbon-rich mineral soils to minimize SOC losses. PMID:26217000

Conversion of lowland tropical forests to tree cash crop plantations loses up to one-half of stored soil organic carbon.

PubMed

van Straaten, Oliver; Corre, Marife D; Wolf, Katrin; Tchienkoua, Martin; Cuellar, Eloy; Matthews, Robin B; Veldkamp, Edzo

2015-08-11

Tropical deforestation for the establishment of tree cash crop plantations causes significant alterations to soil organic carbon (SOC) dynamics. Despite this recognition, the current Intergovernmental Panel on Climate Change (IPCC) tier 1 method has a SOC change factor of 1 (no SOC loss) for conversion of forests to perennial tree crops, because of scarcity of SOC data. In this pantropic study, conducted in active deforestation regions of Indonesia, Cameroon, and Peru, we quantified the impact of forest conversion to oil palm (Elaeis guineensis), rubber (Hevea brasiliensis), and cacao (Theobroma cacao) agroforestry plantations on SOC stocks within 3-m depth in deeply weathered mineral soils. We also investigated the underlying biophysical controls regulating SOC stock changes. Using a space-for-time substitution approach, we compared SOC stocks from paired forests (n = 32) and adjacent plantations (n = 54). Our study showed that deforestation for tree plantations decreased SOC stocks by up to 50%. The key variable that predicted SOC changes across plantations was the amount of SOC present in the forest before conversion--the higher the initial SOC, the higher the loss. Decreases in SOC stocks were most pronounced in the topsoil, although older plantations showed considerable SOC losses below 1-m depth. Our results suggest that (i) the IPCC tier 1 method should be revised from its current SOC change factor of 1 to 0.6 ± 0.1 for oil palm and cacao agroforestry plantations and 0.8 ± 0.3 for rubber plantations in the humid tropics; and (ii) land use management policies should protect natural forests on carbon-rich mineral soils to minimize SOC losses.

Pervasive effects of a dominant foliar endophytic fungus on host genetic and phenotypic expression in a tropical tree

PubMed Central

Mejía, Luis C.; Herre, Edward A.; Sparks, Jed P.; Winter, Klaus; García, Milton N.; Van Bael, Sunshine A.; Stitt, Joseph; Shi, Zi; Zhang, Yufan; Guiltinan, Mark J.; Maximova, Siela N.

2014-01-01

It is increasingly recognized that macro-organisms (corals, insects, plants, vertebrates) consist of both host tissues and multiple microbial symbionts that play essential roles in their host's ecological and evolutionary success. Consequently, identifying benefits and costs of symbioses, as well as mechanisms underlying them are research priorities. All plants surveyed under natural conditions harbor foliar endophytic fungi (FEF) in their leaf tissues, often at high densities. Despite producing no visible effects on their hosts, experiments have nonetheless shown that FEF reduce pathogen and herbivore damage. Here, combining results from three genomic, and two physiological experiments, we demonstrate pervasive genetic and phenotypic effects of the apparently asymptomatic endophytes on their hosts. Specifically, inoculation of endophyte-free (E−) Theobroma cacao leaves with Colletotrichum tropicale (E+), the dominant FEF species in healthy T. cacao, induces consistent changes in the expression of hundreds of host genes, including many with known defensive functions. Further, E+ plants exhibited increased lignin and cellulose content, reduced maximum rates of photosynthesis (Amax), and enrichment of nitrogen-15 and carbon-13 isotopes. These phenotypic changes observed in E+ plants correspond to changes in expression of specific functional genes in related pathways. Moreover, a cacao gene (Tc00g04254) highly up-regulated by C. tropicale also confers resistance to pathogen damage in the absence of endophytes or their products in host tissues. Thus, the benefits of increased pathogen resistance in E+ plants are derived in part from up-regulation of intrinsic host defense responses, and appear to be offset by potential costs including reduced photosynthesis, altered host nitrogen metabolism, and endophyte heterotrophy of host tissues. Similar effects are likely in most plant-endophyte interactions, and should be recognized in the design and interpretation of genetic and phenotypic studies of plants. PMID:25309519

Characterization of the Proteome of Theobroma cacao Beans by Nano-UHPLC-ESI MS/MS.

PubMed

Scollo, Emanuele; Neville, David; Oruna-Concha, M Jose; Trotin, Martine; Cramer, Rainer

2018-02-01

Cocoa seed storage proteins play an important role in flavour development as aroma precursors are formed from their degradation during fermentation. Major proteins in the beans of Theobroma cacao are the storage proteins belonging to the vicilin and albumin classes. Although both these classes of proteins have been extensively characterized, there is still limited information on the expression and abundance of other proteins present in cocoa beans. This work is the first attempt to characterize the whole cocoa bean proteome by nano-UHPLC-ESI MS/MS analysis using tryptic digests of cocoa bean protein extracts. The results of this analysis show that >1000 proteins could be identified using a species-specific Theobroma cacao database. The majority of the identified proteins were involved with metabolism and energy. Additionally, a significant number of the identified proteins were linked to protein synthesis and processing. Several proteins were also involved with plant response to stress conditions and defence. Albumin and vicilin storage proteins showed the highest intensity values among all detected proteins, although only seven entries were identified as storage proteins. A comparison of MS/MS data searches carried out against larger non-specific databases confirmed that using a species-specific database can increase the number of identified proteins, and at the same time reduce the number of false positives. The results of this work will be useful in developing tools that can allow the comparison of the proteomic profile of cocoa beans from different genotypes and geographic origins. Data are available via ProteomeXchange with identifier PXD005586. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sequencing of a QTL-rich region of the Theobroma cacao genome using pooled BACs and the identification of trait specific candidate genes

PubMed Central

2011-01-01

Background BAC-based physical maps provide for sequencing across an entire genome or a selected sub-genomic region of biological interest. Such a region can be approached with next-generation whole-genome sequencing and assembly as if it were an independent small genome. Using the minimum tiling path as a guide, specific BAC clones representing the prioritized genomic interval are selected, pooled, and used to prepare a sequencing library. Results This pooled BAC approach was taken to sequence and assemble a QTL-rich region, of ~3 Mbp and represented by twenty-seven BACs, on linkage group 5 of the Theobroma cacao cv. Matina 1-6 genome. Using various mixtures of read coverages from paired-end and linear 454 libraries, multiple assemblies of varied quality were generated. Quality was assessed by comparing the assembly of 454 reads with a subset of ten BACs individually sequenced and assembled using Sanger reads. A mixture of reads optimal for assembly was identified. We found, furthermore, that a quality assembly suitable for serving as a reference genome template could be obtained even with a reduced depth of sequencing coverage. Annotation of the resulting assembly revealed several genes potentially responsible for three T. cacao traits: black pod disease resistance, bean shape index, and pod weight. Conclusions Our results, as with other pooled BAC sequencing reports, suggest that pooling portions of a minimum tiling path derived from a BAC-based physical map is an effective method to target sub-genomic regions for sequencing. While we focused on a single QTL region, other QTL regions of importance could be similarly sequenced allowing for biological discovery to take place before a high quality whole-genome assembly is completed. PMID:21794110

Sequencing of a QTL-rich region of the Theobroma cacao genome using pooled BACs and the identification of trait specific candidate genes.

PubMed

Feltus, Frank A; Saski, Christopher A; Mockaitis, Keithanne; Haiminen, Niina; Parida, Laxmi; Smith, Zachary; Ford, James; Staton, Margaret E; Ficklin, Stephen P; Blackmon, Barbara P; Cheng, Chun-Huai; Schnell, Raymond J; Kuhn, David N; Motamayor, Juan-Carlos

2011-07-27

BAC-based physical maps provide for sequencing across an entire genome or a selected sub-genomic region of biological interest. Such a region can be approached with next-generation whole-genome sequencing and assembly as if it were an independent small genome. Using the minimum tiling path as a guide, specific BAC clones representing the prioritized genomic interval are selected, pooled, and used to prepare a sequencing library. This pooled BAC approach was taken to sequence and assemble a QTL-rich region, of ~3 Mbp and represented by twenty-seven BACs, on linkage group 5 of the Theobroma cacao cv. Matina 1-6 genome. Using various mixtures of read coverages from paired-end and linear 454 libraries, multiple assemblies of varied quality were generated. Quality was assessed by comparing the assembly of 454 reads with a subset of ten BACs individually sequenced and assembled using Sanger reads. A mixture of reads optimal for assembly was identified. We found, furthermore, that a quality assembly suitable for serving as a reference genome template could be obtained even with a reduced depth of sequencing coverage. Annotation of the resulting assembly revealed several genes potentially responsible for three T. cacao traits: black pod disease resistance, bean shape index, and pod weight. Our results, as with other pooled BAC sequencing reports, suggest that pooling portions of a minimum tiling path derived from a BAC-based physical map is an effective method to target sub-genomic regions for sequencing. While we focused on a single QTL region, other QTL regions of importance could be similarly sequenced allowing for biological discovery to take place before a high quality whole-genome assembly is completed.

Cadmium and cadmium-tolerant soil bacteria in cacao crops from northeastern Colombia.

PubMed

Bravo, D; Pardo-Díaz, S; Benavides-Erazo, J; Rengifo-Estrada, G; Braissant, O; Leon-Moreno, C

2018-05-01

This research aims to assess total-cadmium soil content and microbiological aspects to understand the dynamics of culturable cadmium-tolerant bacteria (CdtB) in cacao soils from northeastern Colombia. An integration of inverted dish plating, Cd determination and a microcalorimetry assay (IMC) was carried out. A farm in Boyacá showed the highest level of total soil Cd (3·74 mg kg -1 ) followed by farms in Santander and Arauca (2·76 and 1·16 mg kg -1 , respectively). Coefficient of determination between total soil Cd and CFU of CdtB was high (R 2  = 0·83) for the farm in Boyacá. Moreover, a pool of 129 CdtB was isolated, and phylogeny of 21 CdtB was discussed. Among CdtB strains isolated, Enterobacter sp. CdDB41 showed major Cd immobilization capacity (Q max of 2·21 and 2·32 J at 6 and 24 mg l -1 of CdCl 2 ), with an immobilization rate of 0·220 mg kg -1  h -1 . Among CdtB strains isolated, Enterobacter sp. CdDB41 showed major Cd immobilization capacity (Q max of 2·21 and 2·32 J at 6 and 24 mg l -1 of CdCl 2 ), with an immobilization rate of 0·220 mg kg -1  h -1 . Nothing is known about soil CdtB in cacao. Our data showed that CdtB such as Enterobacter sp. has high immobilization capacity. Furthermore, the otavite found in situ might be mineralized due to the bacterial metabolic activity of CdtB. © 2018 The Society for Applied Microbiology.

Genomics meets applied ecology: Characterizing habitat quality for sloths in a tropical agroecosystem.

PubMed

Fountain, Emily D; Kang, Jung Koo; Tempel, Douglas J; Palsbøll, Per J; Pauli, Jonathan N; Zachariah Peery, M

2018-01-01

Understanding how habitat quality in heterogeneous landscapes governs the distribution and fitness of individuals is a fundamental aspect of ecology. While mean individual fitness is generally considered a key to assessing habitat quality, a comprehensive understanding of habitat quality in heterogeneous landscapes requires estimates of dispersal rates among habitat types. The increasing accessibility of genomic approaches, combined with field-based demographic methods, provides novel opportunities for incorporating dispersal estimation into assessments of habitat quality. In this study, we integrated genomic kinship approaches with field-based estimates of fitness components and approximate Bayesian computation (ABC) procedures to estimate habitat-specific dispersal rates and characterize habitat quality in two-toed sloths (Choloepus hoffmanni) occurring in a Costa Rican agricultural ecosystem. Field-based observations indicated that birth and survival rates were similar in a sparsely shaded cacao farm and adjacent cattle pasture-forest mosaic. Sloth density was threefold higher in pasture compared with cacao, whereas home range size and overlap were greater in cacao compared with pasture. Dispersal rates were similar between the two habitats, as estimated using ABC procedures applied to the spatial distribution of pairs of related individuals identified using 3,431 single nucleotide polymorphism and 11 microsatellite locus genotypes. Our results indicate that crops produced under a sparse overstorey can, in some cases, constitute lower-quality habitat than pasture-forest mosaics for sloths, perhaps because of differences in food resources or predator communities. Finally, our study demonstrates that integrating field-based demographic approaches with genomic methods can provide a powerful means for characterizing habitat quality for animal populations occurring in heterogeneous landscapes. © 2017 John Wiley & Sons Ltd.

Rejection Thresholds in Chocolate Milk: Evidence for Segmentation

PubMed Central

Harwood, Meriel L.; Ziegler, Gregory R.; Hayes, John E.

2012-01-01

Bitterness is generally considered a negative attribute in food, yet many individuals enjoy some bitterness in products like coffee or chocolate. In chocolate, bitterness arises from naturally occurring alkaloids and phenolics found in cacao. Fermentation and roasting help develop typical chocolate flavor and reduce the intense bitterness of raw cacao by modifying these bitter compounds. As it becomes increasingly common to fortify chocolate with `raw' cacao to increase the amount of healthful phytonutrients, it is important to identify the point at which the concentration of bitter compounds becomes objectionable, even to those who enjoy some bitterness. Classical threshold methods focus on the presence or absence of a sensation rather than acceptability or hedonics. A new alternative, the rejection threshold, was recently described in the literature. Here, we sought to quantify and compare differences in Rejection Thresholds (RjT) and Detection Thresholds (DT) in chocolate milk spiked with a food safe bitterant (sucrose octaacetate). In experiment 1, a series of paired preference tests was used to estimate the RjT for bitterness in chocolate milk. In a new group of participants (experiment 2), we determined the RjT and DT using the forced choice ascending method of limits. In both studies, participants were segmented on the basis of self-declared preference for milk or dark solid chocolate. Based on sigmoid fits of the indifference-preference function, the RjT was ~2.3 times higher for those preferring dark chocolate than the RjT for those preferring milk chocolate in both experiments. In contrast, the DT for both groups was functionally identical, suggesting that differential effects of bitterness on liking of chocolate products are not based on the ability to detect bitterness in these products. PMID:22754143

Rejection Thresholds in Chocolate Milk: Evidence for Segmentation.

PubMed

Harwood, Meriel L; Ziegler, Gregory R; Hayes, John E

2012-10-01

Bitterness is generally considered a negative attribute in food, yet many individuals enjoy some bitterness in products like coffee or chocolate. In chocolate, bitterness arises from naturally occurring alkaloids and phenolics found in cacao. Fermentation and roasting help develop typical chocolate flavor and reduce the intense bitterness of raw cacao by modifying these bitter compounds. As it becomes increasingly common to fortify chocolate with `raw' cacao to increase the amount of healthful phytonutrients, it is important to identify the point at which the concentration of bitter compounds becomes objectionable, even to those who enjoy some bitterness. Classical threshold methods focus on the presence or absence of a sensation rather than acceptability or hedonics. A new alternative, the rejection threshold, was recently described in the literature. Here, we sought to quantify and compare differences in Rejection Thresholds (RjT) and Detection Thresholds (DT) in chocolate milk spiked with a food safe bitterant (sucrose octaacetate). In experiment 1, a series of paired preference tests was used to estimate the RjT for bitterness in chocolate milk. In a new group of participants (experiment 2), we determined the RjT and DT using the forced choice ascending method of limits. In both studies, participants were segmented on the basis of self-declared preference for milk or dark solid chocolate. Based on sigmoid fits of the indifference-preference function, the RjT was ~2.3 times higher for those preferring dark chocolate than the RjT for those preferring milk chocolate in both experiments. In contrast, the DT for both groups was functionally identical, suggesting that differential effects of bitterness on liking of chocolate products are not based on the ability to detect bitterness in these products.

Status of cacao breeding in Ghana

USDA-ARS?s Scientific Manuscript database

Research into cocoa improvement has made a considerable impact on the productivity of the crop in West Africa. Much of the germplasm distributed to farmers have been of Upper Amazon origin following the realisation of their higher agronomic worth over the local Trinitario and Amelonado germplasm. Ho...

Tea, coffee, and cocoa as ultraviolet radiation protectants for beet armyworm nucleopolyhedrovirus

USDA-ARS?s Scientific Manuscript database

The addition of 1% (wt/v) aqueous extracts of cocoa (Theobroma cacao L.) (Malvales: Malvaceae), coffee (Coffea arabica L.) (Gentianales: Rubiaceae), green, and black tea (Camellia sinensis L.) (Ericales: Theaceae) provided excellent ultraviolet (UV) radiation protection for the beet armyworm, Spodo...

Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos by vitrification.

PubMed

Adu-Gyamfi, Raphael; Wetten, Andy

2012-01-01

Losses of cultivated cocoa (Theobroma cacao L.) due to diseases and continued depletion of forests that harbour the wild progenitors of the crop make ex situ conservation of cocoa germplasm of paramount importance. In order to enhance security of in situ germplasm collections, 2-3 mm floral-derived secondary somatic embryos were cryopreserved by vitrification. This work demonstrates the most uncomplicated clonal cocoa cryopreservation. Optimal post-cryostorage survival (74.5 percent) was achieved by 5 d preculture of SSEs on 0.5 M sucrose medium followed by 60 min dehydration in cold PVS2. To minimise free radical related cryo-injury, cation sources were removed from the embryo development solution and/or the recovery medium, the former treatment resulting in a significant benefit. After optimisation with cocoa genotype AMAZ 15, the same protocol was effective across all five additional cocoa genotypes tested. For the multiplication of clones, embryos regenerated following cryopreservation were used as explant sources, and vitrification was found to maintain their embryogenic potential.

Genomic analyses and expression evaluation of thaumatin-like gene family in the cacao fungal pathogen Moniliophthora perniciosa.

PubMed

Franco, Sulamita de Freitas; Baroni, Renata Moro; Carazzolle, Marcelo Falsarella; Teixeira, Paulo José Pereira Lima; Reis, Osvaldo; Pereira, Gonçalo Amarante Guimarães; Mondego, Jorge Maurício Costa

2015-10-30

Thaumatin-like proteins (TLPs) are found in diverse eukaryotes. Plant TLPs, known as Pathogenicity Related Protein (PR-5), are considered fungal inhibitors. However, genes encoding TLPs are frequently found in fungal genomes. In this work, we have identified that Moniliophthora perniciosa, a basidiomycete pathogen that causes the Witches' Broom Disease (WBD) of cacao, presents thirteen putative TLPs from which four are expressed during WBD progression. One of them is similar to small TLPs, which are present in phytopathogenic basidiomycete, such as wheat stem rust fungus Puccinia graminis. Fungi genomes annotation and phylogenetic data revealed a larger number of TLPs in basidiomycetes when comparing with ascomycetes, suggesting that these proteins could be involved in specific traits of mushroom-forming species. Based on the present data, we discuss the contribution of TLPs in the combat against fungal competitors and hypothesize a role of these proteins in M. perniciosa pathogenicity. Copyright © 2015 Elsevier Inc. All rights reserved.

Effects of polyphenol substances derived from Theobroma cacao on gastric mucosal lesion induced by ethanol.

PubMed

Osakabe, N; Sanbongi, C; Yamagishi, M; Takizawa, T; Osawa, T

1998-08-01

The antiulcer activity of cacao liquor water-soluble crude polyphenols (CWSP) was examined. CWSP, alpha-tocopherol, sucralfate (500 mg/kg), and cimetidine (250 mg/kg) were orally administered to male SD rats 30 minutes before ethanol treatment. 5 ml/kg of ethanol given intragastrically caused lesions in mucosa of the glandular stomach. CWSP caused a reduction of such hemorrhagic lesions as well as cimetidine and sucralfate which are typical antiulcer drugs, but alpha-tocopherol was less effective. Thiobarbituric acid reactive substances in gastric mucosa significantly increased with ethanol administration. CWSP treatment significantly reduced this change. The administration of ethanol extensively increased myeloperoxidase (MPO) but not xanthine oxidase (XOD) activity. CWSP reduced the activities of both enzymes; they were considered the main sources of oxygen radicals. According to an in vitro study, CWSP directly reducted XOD but not MPO. These results suggest that the antiulcer mechanism of CWSP was not only radical scavenging but also modulation of leukocyte function.

Impact of condensed tannin size as individual and mixed polymers on bovine serum albumin precipitation.

PubMed

Harbertson, James F; Kilmister, Rachel L; Kelm, Mark A; Downey, Mark O

2014-10-01

Condensed tannins composed of epicatechin from monomer to octamer were isolated from cacao (Theobroma cacao, L.) seeds and added to bovine serum albumin (BSA) individually and combined as mixtures. When added to excess BSA the amount of tannin precipitated increased with tannin size. The amount of tannin required to precipitate BSA varied among the polymers with the trimer requiring the most to precipitate BSA (1000 μg) and octamer the least (50 μg). The efficacy of condensed tannins for protein precipitation increased with increased degree of polymerisation (or size) from trimers to octamers (monomers and dimers did not precipitate BSA), while mixtures of two sizes primarily had an additive effect. This study demonstrates that astringent perception is likely to increase with increasing polymer size. Further research to expand our understanding of astringent perception and its correlation with protein precipitation would benefit from sensory analysis of condensed tannins across a range of polymer sizes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Anti-cariogenic properties of a water-soluble extract from cacao.

PubMed

Ito, Kyoko; Nakamura, Yuko; Tokunaga, Takahisa; Iijima, Daisuke; Fukushima, Kazuo

2003-12-01

The addition of a water-soluble extract from cacao-extracted powder (CEPWS) to a cariogenic model food, a white chocolate-like diet that contains 35% sucrose, significantly reduced caries scores in SPF rats infected with Streptococcus sobrinus 6715, compared to control rats fed a white chocolate-like diet. CEPWS markedly inhibited water-insoluble glucan (WIG) synthesis through crude glucosyltransferases (GTFs) from Streptococcus sobrinus B13N in vitro. GTF-inhibitor(s) in CEPWS was prepared through three-step fractionation, and was termed CEPWS-BT, which is a high molecular weight (>10 kDa) heat-stable matrix of sugar, protein, and polyphenol. When the inhibitory effect of CEPWS-BT on glucan synthesis was examined using the purified GTF-I, GTF-T, and GTF-U enzymes from S. sobrinus B13N, significant reduction in GTF-I and GTF-T activity as a result of adding CEPWS-BT at low concentrations was observed. These results suggest that the addition of CEPWS to cariogenic food could be useful in controlling dental caries.

Geographic List of Prime Contract Awards. Oct 1992-Sep 1993. FY 1993. (Aua, American Samoa-Classified Location Foreign). Part 14

DTIC Science & Technology

1994-03-01

4 en cn aqr . IV qt go 4m Ln co co id rý w It 01)(0 111 Ln cacao 11 cj 4 MW MV." 11 1 ca 0 CD (D C4 cn v (D -4 4 In to U) to W) m (D cn (D -4 w...CY (m C-i In = Go 0 1 90000 11 CL q1t w CI I% cL In CL CL INA CL CL 0) CL W CIL CIL In V CIL ý4 en 11 1 cacao 11 47) 1w In 0) 11 0 1 CIO 0 co If...U)--0)O4 ON ON 3 11 a CONO IICONC 0 00 U> N 0-4 cc- (nWO De u 0 00 ;m -00 F- 00 L 00 .-11J ORICON BI K I0 (1 ý u z a ) 0-00 a- L 0 1 ɘ 10 0.w qt4

In vitro seed germination and rootstock establishing for micrografting of Theobroma cacao L

USDA-ARS?s Scientific Manuscript database

Micrografting has been successfully implemented in several plant species of Acacia, Citrus, Eucalyptus, Havea, Malus, Olea, Opuntia, Prunus and other genera. This technique is employed for plant rejuvenation, true-to-type propagation, genetic improvement, recovery of virus-free plants, testing of po...

The tropical germplasm repository program at the USDA-ARS, Tropical Agriculture Research Station, Mayaguez, Puerto Rico

USDA-ARS?s Scientific Manuscript database

The USDA-ARS Tropical Agriculture Research Station is the only research entity within the National Plant Germplasm system in the insular Caribbean region. It houses germplasm collections of cultivated tropical/subtropical germplasm of bananas/plantains, cacao, mamey sapote, sapodilla, Spanish lime,...

The need for an assembly pilot project

USDA-ARS?s Scientific Manuscript database

Progress has been rapid since the June 2008 start of the cacao genome sequencing project with the completion of the physical map and the accumulation of approximately 10x coverage of the genome with Titanium 454 sequence data of Matina1-6, the highly homozygous Amelonado tree chosen for the project....

21 CFR 163.130 - Milk chocolate.

Code of Federal Regulations, 2010 CFR

2010-04-01

... used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners; (3) Spices, natural and artificial... name of the specific neutralizing agent used in the food. (3) When one or more of the spices... label shall bear an appropriate statement, e.g., “Spice added”, “Flavored with ___”, or “With ___ added...

Cacau Cabruca Agroforestry System of Production in Bahia, Brazil

USDA-ARS?s Scientific Manuscript database

The Cacao Cabruca Agroforestry system of production was developed by farmers in Southern Bahia probably in the beginning of the 19th century. To establish such system, farmers in the Atlantic rain forest region selectively maintained around 75 adult individual native trees per hectare, removed the o...

40 CFR 180.225 - Phosphine; tolerances for residues.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 0.01 Cabbage, Chinese, napa 0.01 Cacao bean, dried bean 0.1 Cashew 0.1 Citron, citrus 0.01 Coffee... Lettuce 0.01 Lime 0.01 Mango 0.01 Millet, grain 0.1 Mushroom 0.01 Nut, brazil 0.1 Oat, grain 0.1 Okra 0.01...

40 CFR 180.225 - Phosphine; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 0.01 Cabbage, Chinese, napa 0.01 Cacao bean, dried bean 0.1 Cashew 0.1 Citron, citrus 0.01 Coffee... Lettuce 0.01 Lime 0.01 Mango 0.01 Millet, grain 0.1 Mushroom 0.01 Nut, brazil 0.1 Oat, grain 0.1 Okra 0.01...

40 CFR 180.225 - Phosphine; tolerances for residues.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 0.01 Cabbage, Chinese, napa 0.01 Cacao bean, dried bean 0.1 Cashew 0.1 Citron, citrus 0.01 Coffee... Lettuce 0.01 Lime 0.01 Mango 0.01 Millet, grain 0.1 Mushroom 0.01 Nut, brazil 0.1 Oat, grain 0.1 Okra 0.01...

40 CFR 180.225 - Phosphine; tolerances for residues.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 0.01 Cabbage, Chinese, napa 0.01 Cacao bean, dried bean 0.1 Cashew 0.1 Citron, citrus 0.01 Coffee... Lettuce 0.01 Lime 0.01 Mango 0.01 Millet, grain 0.1 Mushroom 0.01 Nut, brazil 0.1 Oat, grain 0.1 Okra 0.01...

40 CFR 180.225 - Phosphine; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 0.01 Cabbage, Chinese, napa 0.01 Cacao bean, dried bean 0.1 Cashew 0.1 Citron, citrus 0.01 Coffee... Lettuce 0.01 Lime 0.01 Mango 0.01 Millet, grain 0.1 Mushroom 0.01 Nut, brazil 0.1 Oat, grain 0.1 Okra 0.01...

Wintering ovenbird from Belize recovered on Pennsylvania breeding ground

USGS Publications Warehouse

Dowell, B.A.; Robbins, C.S.

1998-01-01

A first-winter Ovenbird (Seiurus aurocapillus) banded in a cacao plantation in central Belize on 8 February 1989 was found dead on the deck of a home on its breeding ground in northwestern Pennsylvania on 28 June 1997, tying the 9-year age record for the species.

iXora: Haplotype Inferencing and Trait Association

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the source of cocoa beans for chocolate, is an important tropical agriculture commodity that is affected by a number of fungal pathogens and insect pests, as well as concerns about yield and quality. We are trying to find molecular genetic markers that are linked to disease resista...

The Role Of Haplotyping In Mapping The Pod Color Trait

USDA-ARS?s Scientific Manuscript database

Theobroma cacao, the source of cocoa beans for chocolate, is an important tropical agriculture commodity that is affected by a number of fungal pathogens and insect pests, as well as concerns about yield and quality. We are trying to find molecular genetic markers that are linked to disease resista...

The hemibiotrophic cacao pathogen Moniliophthora perniciosa depends on a mitochondrial alternative oxidase for biotrophic development

USDA-ARS?s Scientific Manuscript database

The mitochondrial alternative oxidase (AOX) is a non-energy conserving ubiquinol oxidase found in most fungal genomes studied to date. With the development of fungicides containing cytochrome-dependent respiratory chain (CRC) inhibitors, a strong interest in studying AOX functions in phytopathogenic...

21 CFR 163.130 - Milk chocolate.

Code of Federal Regulations, 2011 CFR

2011-04-01

... used: (1) Cacao fat; (2) Nutritive carbohydrate sweeteners; (3) Spices, natural and artificial... name of the specific neutralizing agent used in the food. (3) When one or more of the spices... label shall bear an appropriate statement, e.g., “Spice added”, “Flavored with ___”, or “With ___ added...

21 CFR 163.117 - Cocoa with dioctyl sodium sulfosuccinate for manufacturing.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Cocoa with dioctyl sodium sulfosuccinate for manufacturing. 163.117 Section 163.117 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION CACAO PRODUCTS Requirements for Specific...

Mapping of quantitative trait loci for butter content and hardness in cocoa beans (Theobroma cacao L.)

USDA-ARS?s Scientific Manuscript database

Cocoa butter is an important raw material for the chocolate, pharmaceutical, and cosmetic industries. The butter content and quality in cocoa beans are genetically controlled characteristics, and affect its commercial values and industrial applicability. In the present work, an F2 population derived...

Pooled-BAC sequencing of a black pod resistance region (cBPQTL12) in T. cacao

USDA-ARS?s Scientific Manuscript database

Whole genome sequencing (WGS) is an expensive and technically challenging endeavor. An alternative to WGS is to sequence specific chromosomal segments of biological interest (e.g. a QTL interval). This method is cheaper than WGS and reduces the risk of misassembly from distal parts of the genome. Us...

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

USDA-ARS?s Scientific Manuscript database

The International Cocoa Genebank, Trinidad (ICG,T) is the largest public domain field gene bank collection of cacao 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 1480 t...

Integrating genomics into future approaches for cocoa selection and propagation in Côte d’Ivoire

USDA-ARS?s Scientific Manuscript database

In Côte-d’Ivoire cocoa breeding is based on a reciprocal recurrent scheme that has been prepared with the aim of improving simultaneously the characteristics of the two main populations: Upper Amazon and Lower Amazon+ Trinitario. Resistance to Phytophtora and Cacao Swollen Shoot Virus has become th...

Evaluation of soil quality in areas of cocoa cabruca, forest and multicropping in southern Bahia, Brazil

USDA-ARS?s Scientific Manuscript database

The Atlantic Rain Forest is one of the most complex natural environments of the earth and, linked with this ecosystem, the cacao-cabruca system is agroforestry cultivation with an arrangement including a range of environmental, social and economical benefits and can protect many features of the biod...

21 CFR 163.155 - Milk chocolate and vegetable fat coating.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Milk chocolate and vegetable fat coating. 163.155... § 163.155 Milk chocolate and vegetable fat coating. (a) Description. Milk chocolate and vegetable fat...) Safe and suitable vegetable derived oils, fats, and stearins other than cacao fat. The oils, fats, and...

21 CFR 163.155 - Milk chocolate and vegetable fat coating.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Milk chocolate and vegetable fat coating. 163.155... § 163.155 Milk chocolate and vegetable fat coating. (a) Description. Milk chocolate and vegetable fat...) Safe and suitable vegetable derived oils, fats, and stearins other than cacao fat. The oils, fats, and...

21 CFR 163.155 - Milk chocolate and vegetable fat coating.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Milk chocolate and vegetable fat coating. 163.155... § 163.155 Milk chocolate and vegetable fat coating. (a) Description. Milk chocolate and vegetable fat...) Safe and suitable vegetable derived oils, fats, and stearins other than cacao fat. The oils, fats, and...

On the importance of the Cerulean and Golden-winged Warblers summits in the National Federation of Coffee Growers of Colombia in Bogotá

Treesearch

Paul B. Hamel

2008-01-01

Cerulean Warbler is a bird with problems; this migratorybird lives in environments on which large numbersof people depend for an adequate productivelivelihood, energy, high quality wood products, coffee,and cacao. Solving the biological problems of thisspecies in its complex...

Discovery and mapping of a new expressed sequence tag-single nucleotide polymorphism and simple sequence repeat panel for large-scale genetic studies and breeding of Theobroma cacao L.

PubMed Central

Allegre, Mathilde; Argout, Xavier; Boccara, Michel; Fouet, Olivier; Roguet, Yolande; Bérard, Aurélie; Thévenin, Jean Marc; Chauveau, Aurélie; Rivallan, Ronan; Clement, Didier; Courtois, Brigitte; Gramacho, Karina; Boland-Augé, Anne; Tahi, Mathias; Umaharan, Pathmanathan; Brunel, Dominique; Lanaud, Claire

2012-01-01

Theobroma cacao is an economically important tree of several tropical countries. Its genetic improvement is essential to provide protection against major diseases and improve chocolate quality. We discovered and mapped new expressed sequence tag-single nucleotide polymorphism (EST-SNP) and simple sequence repeat (SSR) markers and constructed a high-density genetic map. By screening 149 650 ESTs, 5246 SNPs were detected in silico, of which 1536 corresponded to genes with a putative function, while 851 had a clear polymorphic pattern across a collection of genetic resources. In addition, 409 new SSR markers were detected on the Criollo genome. Lastly, 681 new EST-SNPs and 163 new SSRs were added to the pre-existing 418 co-dominant markers to construct a large consensus genetic map. This high-density map and the set of new genetic markers identified in this study are a milestone in cocoa genomics and for marker-assisted breeding. The data are available at http://tropgenedb.cirad.fr. PMID:22210604

Independent origins and incipient speciation among host-associated populations of Thielaviopsis ethacetica in Cameroon.

PubMed

Mbenoun, Michael; Wingfield, Michael J; Letsoalo, Teboho; Bihon, Wubetu; Wingfield, Brenda D; Roux, Jolanda

2015-11-01

Thielaviopsis ethacetica was recently reinstated as a distinct taxon using DNA phylogenies. It is widespread affecting several crop plants of global economic importance. In this study, microsatellite markers were developed and used in conjunction with sequence data to investigate the genetic diversity and structure of Th. ethacetica in Cameroon. A collection of 71 isolates from cacao, oil palm, and pineapple, supplemented with nine isolates from other countries were analysed. Four genetic groups were identified. Two of these were associated with oil palm in Cameroon and showed high genetic diversity, suggesting that they might represent an indigenous population of the pathogen. In contrast, the remaining two groups, associated with cacao and pineapple, had low genetic diversity and, most likely, represent introduced populations. There was no evidence of gene flow between these groups. Phylogenetic analyses based on sequences of the tef1-α as well as the combined flanking regions of six microsatellite loci were consistent with population genetic analyses and suggested that Th. ethacetica is comprised of two divergent genetic lineages. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Phylogenetic analysis of DNA and RNA polymerases from a Moniliophthora perniciosa mitochondrial plasmid reveals probable lateral gene transfer.

PubMed

Andrade, B S; Góes-Neto, A

2015-10-30

The filamentous fungus Moniliophthora perniciosa is a hemibiotrophic basidiomycete that causes witches' broom disease of cacao (Theobroma cacao L.). Many fungal mitochondrial plasmids are DNA and RNA polymerase-encoding invertrons with terminal inverted repeats and 5'-linked proteins. The aim of this study was to carry out comparative and phylogenetic analyses of DNA and RNA polymerases for all known linear mitochondrial plasmids in fungi. We performed these analyses at both gene and protein levels and assessed differences between fungal and viral polymerases in order to test the lateral gene transfer (LGT) hypothesis. We analyzed all mitochondrial plasmids of the invertron type within the fungal clade, including five from Ascomycota, seven from Basidiomycota, and one from Chytridiomycota. All phylogenetic analyses generated similar tree topologies regardless of the methods and datasets used. It is likely that DNA and RNA polymerase genes were inserted into the mitochondrial genomes of the 13 fungal species examined in our study as a result of different LGT events. These findings are important for a better understanding of the evolutionary relationships between fungal mitochondrial plasmids.

Anti-inflammatory properties of clovamide and Theobroma cacao phenolic extracts in human monocytes: evaluation of respiratory burst, cytokine release, NF-κB activation, and PPARγ modulation.

PubMed

Zeng, Huawu; Locatelli, Monica; Bardelli, Claudio; Amoruso, Angela; Coisson, Jean Daniel; Travaglia, Fabiano; Arlorio, Marco; Brunelleschi, Sandra

2011-05-25

There is a great interest in the potential health benefits of biologically active phenolic compounds in cocoa (Theobroma cacao) and dark chocolate. We investigated the anti-inflammatory potential of clovamide (a N-phenylpropenoyl-L-amino acid amide present in cocoa beans) and two phenolic extracts from unroasted and roasted cocoa beans, by evaluating superoxide anion (O(2)(-)) production, cytokine release, and NF-κB activation in human monocytes stimulated by phorbol 12-myristate 13-acetate (PMA). The effects of rosmarinic acid are shown for comparison. Clovamide and rosmarinic acid inhibited PMA-induced O(2)(-) production and cytokine release (with a bell-shaped curve and maximal inhibition at 10-100 nM), as well as PMA-induced NF-κB activation; the two cocoa extracts were less effective. In all tests, clovamide was the most potent compound and also enhanced peroxisome proliferator-activated receptor-γ (PPARγ) activity, which may exert anti-inflammatory effects. These findings indicate clovamide as a possible bioactive compound with anti-inflammatory activity in human cells.

Patterns of chocolate consumption.

PubMed

Seligson, F H; Krummel, D A; Apgar, J L

1994-12-01

Although consumed in some form since at least 460 AD, cacao (Theobroma cacao) was not used in confectionery until the 19th century when the cocoa press was invented. Per capita consumption of chocolate confectionery in the United States is moderate (approximately 4.6-4.8 kg/y) compared with that of many northern European countries (approximately 7-10 kg/y). Eleven percent of the US population reported consuming chocolate candy on > or = 1 of the 3 d of recorded food intake in the US Department of Agriculture Nationwide Food Consumption Survey 1987-1988; < 1.0% consumed chocolate every day. The Western region of the United States contained the highest proportion of chocolate consumers. More whites than other racial groups were consumers. Chocolate was consumed by more people in the winter than in other seasons and more was consumed at snacks than at meals. The mean amount of chocolate consumed was approximately 30-90 g/d, depending on sex and age group. Chocolate candy was only a minor contributor (0.7-3.4%) to the overall dietary intake of total energy, fat, saturated fatty acids, and stearic acid.

Anti-Oxidative Polyphenolic Compounds of Cocoa.

PubMed

Nabavi, Seyed F; Sureda, Antoni; Daglia, Maria; Rezaei, Parizad; Nabavi, Seyed M

2015-01-01

Oxidative stress plays a key role in the pathogenesis of different serious chronic diseases such as cancer, diabetes, cardiovascular and neurodegenerative disorders, etc. Recent research has been focused on the beneficial role of dietary antioxidants against oxidative stress both under in vitro and in vivo conditions. Theobroma cacao L. (cacao tree) is an evergreen tree which is native to South America. It is a plant of great economic importance and its seeds are commonly used to produce cocoa powder and chocolate. In addition to its uses in food industry, cocoa is a rich source of polyphenolic antioxidants. There is a plethora of in vitro and in vivo studies that report cocoa antioxidant capacity. The protective activity of cocoa seems to be due to its phytochemical constituents, especially catechins. However, bioavailability of cocoa polyphenolic constituents following oral administration is very low (nanomolar concentrations). In the present paper, we critically reviewed the available literature on the antioxidant and free radical scavenging activities of cocoa and its polyphenolic constituents. In addition to these, we provide brief information about cultivation, phytochemistry, bioavailability and clinical impacts of cocoa.

Application of nuclear energy to agriculture. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moh, C.C.

The following research projects in radiation botany were conducted: mutation breeding of beans and cassava; biological response of coffee plants; and radiosensitivity of tropical plants. In the field of entomology experiments were conducted on radiosterilization of the Mediterranean fruit fly, the coffee leaf miner, the torsalo and the meliaceous shootborer. The following research projects in plant physiology were conducted: physiology of cassava plants; effects of temperature on germination of cacao seeds; physiology of cacao seeds; sulfur metabolism using /sup 35/S; diseases and parasites of banana fruits; the mechanism controlling dwarfism in a radioinduced single gene bean mutant; and the usemore » of wetting agents in foliar nutrition. The following research projects in soil chemistry were conducted: acidity and cation movement in tropical soils; phosphate in soils of the humid tropics; movement, adsorption and desorption of sulfates; free iron and aluminium oxides in tropical soils; mineralization of organic nitrogen in soils on volcanic materials; soil chemical properties of recent volcanic ash; and spatial distribution of the absorbing roots in coffee. Discussions are presented of installation of radiation facilities and collection of rainfall for fallout analysis. (HLW)« less

Performance and Loads Data from a Wind Tunnel Test of a Full-Scale Rotor with Four Blade Tip Planforms

DTIC Science & Technology

1980-09-01

a.c ~~cceccc.~~CCo COeCO @Coc Cacao ace cc* Ow. a 6 -**40 r’M18 - 4 5* 1~ *4 I VII I I cccccccco CeCCCCCC cccoo cacao acccC acC eccc t4 C m 9- VFAFA...0 Q- 0W%0 0 *0 00N’ 0M 00M 0 00 0 00 a. De (A m~) 0010 moo 0 t- 0 N fl,0 tt- 0O t0%0 N-4 co 0 N cc Z 0 -. 4b . N-4 0 F400 4 0 0 0 00 00 a 40 040 .0...C3M C 0 0.-e 00-~ 0 0 LL V4% % De ON WON r’- eN Ml.ý4N ’tGo N Nf Ln N0 0.00 oA𔃺 MNO 4NO ONO x Wr a t.-0 0o a)o ~0 1,- 0 000 000 t-UQ tj n6os f- o 0

Geographic List of Prime Contract Awards. Oct 1992-Sep 1993. FY 1993. (Acton (Sta), Mass.-Wayne County, Missouri). Part 7

DTIC Science & Technology

1994-03-01

ED00LEI00ED00 0D 00C ( o DE O 1 c’m mmlmmm am mm~ mII MeO ds~)(4 - -4,.4-I4 .-4 ’ 4 -4 -4 4 No v0 to M (V) 110 1 0 4’) If I (i9tot i 0 (20) D(1...ooIm~0o Uom4-( el.W it I W00 II’a4- 00 W4al 000 0al0al)04fal -n U )-4 as W IfI cacao IIZN.-4 V-4 &A-4 v -4 to CO-E..4-4 " .*-E4 4 -4 -4U1r -44-v .4 -40...n E 44V4.*4 x Cn 11 cacao MI mn -4.E W CON Off 0 00050 If >- 11 01 cooG it -4 _a LLa 11 c 1 0000 1 N4cr 11 100 0 W 1 11 1 E for 10 -4- -4 -.4 -4- ý4

Alpha List of Prime Contract Awards. Oct 92-Sep 93. FY93. (Food Services INc. - Greenbay Supply Co. Inc.). Part 8

DTIC Science & Technology

1994-03-01

C4 IaN I-INY "-I4..4.4.4 1 4 0 0 0 0 0 0 0 0 IL K m0 I 00.4 0 "-t W In x G 1-0000 i-10100 I-0 00 0 000n qoo ~ U 40 I in0.-4 It -I.. XINz dE £0 Zqt0 V0...14 W 0. I 004’ 1: ONO-40- -4 CU)00 --4 -. 4 0) .- NN 0 0) (flN0-- N-W4V .- 400-Nd0) 4-4- .4 N -4 471 14 I cacao 1O OH I to00No aI .as 411 1: 1 0W N...uI 1 00 0 a) " Z >.Z= 2C sZ >tZZ z Z 2C z CZ Z CZ z cacao o . go• Je 0 c 0 qto .19 Co M• $_ .o0 Su 0 I IŕO 0 00 0 ONO N 0 00 W ! "-C" (a a HI 1000

Morphological, biochemical, molecular and ultrastructural changes induced by Cd toxicity in seedlings of Theobroma cacao L.

PubMed

Castro, Andressa V; de Almeida, Alex-Alan F; Pirovani, Carlos P; Reis, Graciele S M; Almeida, Nicolle M; Mangabeira, Pedro A O

2015-05-01

Seeds from Theobroma cacao progenies derived from the self-pollination of 'Catongo'×'Catongo' and the crossing between CCN-10×SCA-6 were immersed for 24h in different Cd solutions (2; 4; 8; 16 and 32 mgL(-1)) along with the control treatment (without Cd). Shortly after, the seeds were sown in plastic tubes containing organic substrate and were grown in a greenhouse for 60 days. The treatment with Cd was observed to cause morphological, biochemical, molecular and ultrastructural changes in both progenies of T. cacao. There has been deformation in chloroplasts, nuclear chromatin condensation, and reduction in thickness of the mesophyll. As for 'Catongo'×'Catongo', a decrease in thickness of the epidermis was noted on the abaxial face. There has been increased guaiacol peroxidase activity in the roots of CCN-10×SCA-6, as well as in the''Catongo'×'Catongo' leaves. In the presence of Cd, CCN-10×SCA-6 showed increased expression of the genes associated with the biosynthesis of phytochelatin (PCS-1) and class III peroxidases (PER-1) in leaves, and metallothionein (MT2b), in roots. In 'Catongo'×'Catongo', there has been an increase in the expression of genes associated with the biosynthesis of PER-1 and cytosolic superoxide dismutase dependent on copper and zinc (Cu-Zn SODCyt) in leaves and from MT2b and PCS-1 and roots. There was higher accumulation of Cd in the aerial parts of seedlings from both progenies, whereas the most pronounced accumulation was seen in''Catongo'×'Catongo'. The increase in Cd concentration has led to lower Zn and Fe levels in both progenies. Hence, one may conclude that the different survival strategies used by CCN-10×SCA-6 made such progeny more tolerant to Cd stress when compared to''Catongo'×'Catongo'. Copyright © 2015. Published by Elsevier Inc.

Foraging ecology of howler monkeys in a cacao (Theobroma cacao) plantation in Comalcalco, Mexico.

PubMed

Muñoz, David; Estrada, Alejandro; Naranjo, Eduardo; Ochoa, Susana

2006-02-01

Recent evidence indicates that primate populations may persist in neotropical fragmented landscapes by using arboreal agroecosystems, which may provide temporary habitats, increased areas of vegetation, and connectivity, among other benefits. However, limited data are available on how primates are able to sustain themselves in such manmade habitats. We report the results of a 9-month-long investigation of the feeding ecology of a troop of howler monkeys (n = 24) that have lived for the past 25 years in a 12-ha cacao plantation in the lowlands of Tabasco, Mexico. A vegetation census indicated the presence of 630 trees (> or =20 cm diameter at breast height (DBH)) of 32 shade species in the plantation. The howlers used 16 plant species (13 of which were trees) as sources of leaves, fruits, and flowers. Five shade tree species (Ficus cotinifolia, Pithecellobium saman, Gliricidia sepium, F. obtusifolia, and Ficus sp.) accounted for slightly over 80% of the total feeding time and 78% of the total number trees (n = 139) used by the howlers, and were consistently used by the howlers from month to month. The howlers spent an average of 51% of their monthly feeding time exploiting young leaves, 29% exploiting mature fruit, and 20% exploiting flowers and other plant items. Monthly consumption of young leaves varied from 23% to 67%, and monthly consumption of ripe fruit varied from 12% to 64%. Differences in the protein-to-fiber ratio of young vs. mature leaves influenced diet selection by the monkeys. The howlers used 8.3 ha of the plantation area, and on average traveled 388 m per day in each month. The howlers preferred tree species whose contribution to the total tree biomass and density was above average for the shade-tree population in the plantation. Given the right conditions of management and protection, shaded arboreal plantations in fragmented landscapes can sustain segments of howler monkey populations for many decades. Copyright 2006 Wiley-Liss, Inc.

Drought effects on soil carbon dioxide production in two ecosystems in Central Sulawesi, Indonesia

NASA Astrophysics Data System (ADS)

van Straaten, Oliver

2010-05-01

Drought response on soil CO2 production dynamics were examined in two tropical ecosystems in central Sulawesi, Indonesia. Large-scale throughfall displacement roofs were built in a cacao (Theobroma cacao) / Gliricidia sepium agroforestry plantation (560 m.a.s.l.) and in a sub-montane tropical rainforest (1050 m.a.s.l.) to simulate drought conditions. At each site, ecosystem drought responses from three roof plots were compared to three undisturbed control plots. Soil CO2 production was measured spatially at the soil surface and vertically within the soil profile to 2.5 m depth every two weeks. 1. The cacao / Gliricidia ecosystem exhibited a mild drought response. Here, soil CO2 production decreased by 13% in comparison to the control plots during the 13 month induced drought. The mild drought response is attributed to two reasons. First, soil CO2 efflux exhibited an inverse parabolic relationship with soil moisture (R2 = 0.32): soil CO2 efflux peaked at intermediate moisture conditions, but was low when soil conditions became dry (in the induced drought plots), and when the soil became water saturated (in the control plots). This means that respiration differences between control and roof plots may have been masked when soil moisture conditions were saturated in the control and concurrently dry in roof plots. Secondly, the shallow rooted cacao understory grown next to the deeper rooted Gliricidia overstory created a favourable set of site conditions that enabled the ecosystem to mitigate serious drought stress. The experiment had a CO2 neutral effect overall: emissions were initially reduced during the induced drought period but rebounded and surpassed the control during the five month rewetting phase, thus compensating for earlier declines. 2. In contrast, the sub-montane tropical rainforest experienced a severe decrease in soil CO2 production. Here, soil CO2 efflux decreased by an average of 39% in comparison to the control during the 24 month induced drought period. Soil moisture, the main variable controlling CO2, exhibited a strong positive linear relationship with soil CO2 production (R2 = 0.72). A two phase ecosystem drought response was observed. During the first phase, which lasted nine months, leaf litter respiration declined while the total respiration from autotrophic and belowground heterotrophic sources remained relatively unchanged, although an upward shift from the subsoil to the soil surface was measured. During the second phase of the experiment, when drought conditions intensified further (the next 16 months), belowground CO2 production from heterotrophic and autotrophic sources decreased at all soil depths. Leaf litter respiration remained negligible. Recuperation after the drought was slow in this ecosystem and did not rebound to control plot levels. In this ecosystem, the simulated drought resulted in a reduction in overall CO2 emission.

Colletotrichum gloeosporioides s.l. associated with Theobroma cacao and other plants in Panama: multilocus phylogenies distinguish host-associated pathogens from asymptomatic endophytes

USDA-ARS?s Scientific Manuscript database

Species of Colletotrichum interact with a vast but as yet undetermined number of plant species as pathogens and as asymptomatic endophytes. It is not known, however, whether these contrasting ecological modes are optional strategies exercised by individual species or whether species ecology is more ...

Heavy metals in soils of cocoa plantation (Theobroma cacao L.)

USDA-ARS?s Scientific Manuscript database

Cocoa has experienced significant growth in recent years in Peru and the presence of heavy metals in the soils of these plantations is a potential problem for the export of this product. Contents of heavy metals (Cd, Ni, Pb, Fe, Cu, Zn, Mn) in soils from 19 plantations that have been in production f...

The Search for Value and Meaning in the Cocoa Supply Chain in Costa Rica

Treesearch

Jessica Haynes; Frederick Cubbage; D. Evan Mercer; Erin Sills

2012-01-01

Qualitative interviews with participants in the cocoa (Theobroma cacao) supply chain in Costa Rica and the United States were conducted and supplemented with an analysis of the marketing literature to examine the prospects of organic and Fairtrade certification for enhancing environmentally and socially responsible trade of cocoa from Costa Rica. Respondents were...