Entry: Bill Guyton, World Cocoa Foundation
Through the
Cocoa Borlaug Fellowship Program, the U.S. Department of Agriculture/Foreign Agricultural Service and the World Cocoa Foundation have supported the Cocoa Research Institute of Ghana’s breeding program. In 2008, the program supported cocoa breeder Abu Dadzie to complete a fellowship with the U.S. Department of Agriculture/Agricultural Research Service (USDA/ARS) in Miami. The Cocoa Research Institute of Ghana (CRIG) then supported him to continue his stay into mid-2009. In August/September 2009, USDA/ARS Miami supported Kathleen Kelley to travel to Ghana to provide training to CRIG’s laboratory technicians. Below is an excerpt from the report on her work.
A problem facing cacao breeders is the potential for misidentification of the clones used to produce hybrid seedlings through mislabeling or errors in clonal propagation. A recently developed genetic test, the single nucleotide polymorphism (SNP) assay, can identify such mislabeling. Two significant advantages of the SNP assay over the currently more commonly used microsatellite genetic marker assay are that SNPs can be reliably assayed without expensive electrophoretic equipment and that the SNP assay results in an unambiguous genotype requiring no further post-assay analysis as is the case with microsatellites. The SNP assay is a small first step to making reliable genotyping available in cocoa producing countries.
In August 2009, the Cocoa Research Institute of Ghana (CRIG) acquired a fluorescent plate reader and the probes and primers necessary to do SNP assays of cocoa plants. Performing the assays, however, requires some technical expertise. The training provided at the installation of the plate reader consisted of a PowerPoint presentation by the installer on the potential uses of the equipment. As the technicians in the laboratory were unfamiliar with both the equipment and the specialized SNP assay that the equipment was for, additional training was necessary. Dr. David Kuhn, one of the molecular biologists at the USDA-ARS Subtropical Horticulture Research Station (SHRS) lab involved in the cocoa genotyping project, suggested that someone familiar with both the techniques and with the Ghanaian work environment be sent to Ghana to train the laboratory technicians. Kathleen Kelley was chosen for this job. Kathleen is currently a graduate student in Chemistry at the University of Illinois at Urbana-Champaign and recently returned from two years (2006-8) teaching secondary school science in Ghana in the Peace Corps. After being trained in the SNP assay at the USDA station in Miami, FL, Kathleen returned to Ghana for a month to teach the CRIG staff the procedures to quantify the amount of DNA and subsequently run SNP assays on their cocoa samples. The extended stay was designed to assure that the operators became completely comfortable with the theory and operation of the plate reader and the assays that they would be performing, as well as to allow for any delays from equipment malfunction or difficulty in shipping.
DNA Quantification
Over the course of the month, Kathleen trained the permanent technicians at the CRIG Molecular Biology laboratory in two assays. The first was a simple quantification protocol that measured the amount of DNA in an extracted sample by binding a fluorescent SYBRGreen dye to the DNA and comparing the fluorescence to a standard curve. After the practice gained in making a satisfactory standard curve, however, the group had few problems getting decent results in the SYBRGreen assay and figuring the volumes of sample needed to do the polymerase chain reactions (PCR) that are part of the SNP procedure. The CRIG lab anticipates being able to quantify DNA for cocoa and all the other plant species involved in their research.
SNP Assays
The standard laboratory procedure for genetic analysis is a polymerase chain reaction (PCR) of DNA fragments isolated from individuals of interest, followed by agarose gel electrophoresis of the PCR products and comparison of electrophoretic banding patterns among individuals. This procedure can take upwards of an hour and frequently gives uncertain results. The SNP assay in which Kathleen was training the laboratory technicians completely eliminated the agarose gel stage and substituted, instead, a two-minute fluorescence reading on the plate reader before and after PCR. Because the PCR reaction was quite robust and the samples were classified qualitatively according to the differences in their pre-and post-reads, careful mixing and pipetting were not as much of a challenge as they were in the DNA quantification, where the samples were compared to an external standard curve. Problems were encountered when samples failed to amplify enough in the PCR to give a fluorescence signal that allowed the technicians to definitively say whether the sample was homozygous or heterozygous at that allele. However, because the fluorescence read took only two minutes and was completely non-destructive, the solution to this problem was that if a sample failed to amplify, it was simply put through another round of PCR, and the readings on the sample were retaken. This process allowed the technicians to genotype samples that they were not able to assign the first time. This solution would not have been possible with an agarose gel procedure; in addition to the time cost, agarose gels would render the sample useless for any further analysis.
Equipment in DNA analysis lab
Uses for the SNP Assay
The SNP assays are very powerful tools for genotyping individuals. An example of the data that the new assays can now provide CRIG researchers and plant breeders came from samples that the technicians were using for training. They were given five samples of the PA 150 clone included in the CRIG clonal stock. The genotypes for four of the samples of that clone were the same at each SNP tested. One sample, however, was consistently different from the other four. Either this sample had been mislabeled in the collection, or, more seriously, a tree that CRIG was using in its breeding program was not what the breeders believed it to be. Kathleen and the technicians were able to draw this conclusion just by looking at the results of the first test; running subsequent SNP assays that used other probes and primers simply confirmed what they already suspected. In this instance, the assay showed that something was wrong because the unusual sample was compared to other samples in the same sample set that were theoretically genetically identical. As time goes by, however, CRIG will build up a database of SNP genotypes for all of their clones, so they will be able to compare a sample with their records to see if the sample is what they expect it to be. The assay will also be used to identify clones in the seed gardens, and to make sure that crosses have genotypes consistent with the DNA of the parents.
Entry: Bill Guyton, World Cocoa Foundation
In 1986, a consortium of chocolate manufacturing companies and The Pennsylvania State University (PSU) established the Penn State Endowed Program to support improvements in cocoa production through research on Theobroma cacao, the chocolate tree. This program operates at PSU with revenues from an endowment, funded by World Cocoa Foundation member companies. Additional contributions are also made by PSU, and the United States Department of Agriculture (USDA) and other sources.
Over the past decade alone, the program has supported eight Ph.D. graduates and nearly 30 undergraduates. Graduates have gone on to work in government, academics and the private sector. Additionally, the program has welcomed six visiting scientists from cocoa-producing countries over the last four years.
The goals of the Penn State Endowment Program are:
- To stabilize and regionally diversify cocoa production
- To promote increased cocoa production proportional to demand
- To improve economic status of cacao farmers and producing countries
- To protect rainforest habitat and associated species through promotion of sustainable and profitable cacao
production systems
This is accomplished through the identification of elite cacao germplasm, propagation systems and technology transfer to cocoa farmers. The program is under the excellent leadership of Dr. Mark Guiltinan. For more information on the program, please visit: http://guiltinanlab.cas.psu.edu/Research/Cocoa/cocoa.htm.
Penn State Endowment Program faculty and students
I have the opportunity to visit Penn State University each fall, for an annual review of the program. It is encouraging to learn about the accomplishments of the program and the collaborations the PSU team has formed with researchers in cocoa-producing countries. Some of these achievements include the development of vegetative propagation methods that can be used in combination to achieve a large and rapid multiplication of cacao plants from single elite trees including in vitro tissue culture. The PSU program is also helping to identify ways of reducing disease losses on cocoa farms, as highlighted in one of our
past blogs by PSU doctoral graduate, Rachel Melnick.
The World Cocoa Foundation is pleased to support the PSU cocoa research endowment with our member companies and thank Dr. Mark Guiltinan, Siela Maximova, the faculty and students who have made the program a success.
Entry: Robert Peck, World Cocoa Foundation
Last week I had the opportunity to visit the department of Huila in Colombia with Bernardo Saenz (Executive Director of Colombia’s National Cocoa Council), Juan Carlos Arroyave and Alberto Agudelo (from the World Cocoa Foundation’s member company Casa Luker) and Ricardo Mejia (from the World Cocoa Foundation’s member company Compañía Nacional de Chocolates). The World Cocoa Foundation and our two member companies are funding training activities through the National Cocoa Council in this region. I was warned to be prepared to travel to a valley which is suffering from a severe drought and where average temperatures easily surpass 110 F (43 C). Huila, located in the south-west of the country, has traditionally been known for its role as a rice growing region (using flood irrigation), its oil exploration fields, and more recently for aquaculture initiatives and the production of specialty coffees in the higher altitudes of the Andean mountain range. Cocoa has played an important role on a local scale, but poor fermentation practices and the presence of monilia (frosty pod rot) have severely affected yields and farm incomes.
I would like to share with you the story of a farmer I met nearby the municipality of Rivera. Omar is 73 years old and for the past 9 years has lived with his wife on a 4-hectare (9.8-acre) farm. When he purchased the farm, it was generating almost no income and the cocoa trees had been neglected, reaching over 15 feet in height and producing only a few pods per year. Omar has tried growing other crops (tobacco and passion fruit), but low prices and the lack of market opportunities made these not a sustainable option. Omar began receiving technical assistance from Casa Luker two years ago, and his farm has been selected as a demonstration site for training that the World Cocoa Foundation is co-funding on better post-harvest practices. Reducing the crop height, continual pruning, removal of infected pods, grafting and irrigation are some of the practices that he has implemented. What are the results?
Crop Year Cocoa Production Estimated Net Profit
2006-2007 year 330 Kg / ha 646,500 Colombian pesos
2007-2008 year 557 Kg / ha 913, 480 Colombian pesos
2008-2009 year 1,400 Kg / ha 3,872,000 Colombian pesos
Omar is definitely a success story. Due to the amount of work he needs to invest into his farm, he has had to hire neighbors to help him with the agricultural practices that he has adopted. He is fortunate to have enough supply of water on his farm, which in the last year had been critical for his crops to thrive despite the dry season. The incidence of monilia in his crop is nowadays less than 5%, producing a positive effect on his bottom line. I am hopeful that next time I visit Omar, he will have finished grafting his unproductive cocoa trees, his timber trees will be a few feet higher and hopefully his neighbors have also made similar progress because Omar’s success is evident.
Entry: Robert Peck, World Cocoa Foundation
Last week I had the opportunity to visit the department of Huila in Colombia with Bernardo Saenz (Executive Director of Colombia’s National Cocoa Council), Juan Carlos Arroyave and Alberto Agudelo (from the World Cocoa Foundation’s member company Casa Luker) and Ricardo Mejia (from the World Cocoa Foundation’s member company Compañía Nacional de Chocolates). The World Cocoa Foundation and our two member companies are funding training activities through the National Cocoa Council in this region. I was warned to be prepared to travel to a valley which is suffering from a severe drought and where average temperatures easily surpass 110 F (43 C). Huila, located in the south-west of the country, has traditionally been known for its role as a rice growing region (using flood irrigation), its oil exploration fields, and more recently for aquaculture initiatives and the production of specialty coffees in the higher altitudes of the Andean mountain range. Cocoa has played an important role on a local scale, but poor fermentation practices and the presence of monilia (frosty pod rot) have severely affected yields and farm incomes.
Photo: Omar holds a cocoa pod infected with monilia (left) and a healthy pod (right).
I would like to share with you the story of a farmer I met nearby the municipality of Rivera. Omar is 73 years old and for the past 9 years has lived with his wife on a 4-hectare (9.8-acre) farm. When he purchased the farm, it was generating almost no income and the cocoa trees had been neglected, reaching over 15 feet in height and producing only a few pods per year. Omar has tried growing other crops (tobacco and passion fruit), but low prices and the lack of market opportunities made these not a sustainable option. Omar began receiving technical assistance from Casa Luker two years ago, and his farm has been selected as a demonstration site for training that the World Cocoa Foundation is co-funding on better post-harvest practices. Reducing the crop height, continual pruning, removal of infected pods, grafting and irrigation are some of the practices that he has implemented. What were the results?
In 2006-2007, he produced 330 Kg/ha resulting in an estimated net profit of 646,500 Colombian pesos. In the following two years, Omar’s cocoa production continued to increase; in 2007-2008, he produced 557 Kg/ha resulting in an estimated net profit of 913,480 Colombian pesos, and in 2008-2009 he produced 1,400 Kg/ha leading to an estimated net profit of 3,872,000 Colombian pesos.
Photo: A grafted seedling on Omar’s farm.
Omar is definitely a success story. Due to the amount of work he needs to invest into his farm, he has had to hire neighbors to help him with the agricultural practices that he has adopted. He is fortunate to have enough supply of water on his farm, which in the last year had been critical for his crops to thrive despite the dry season. The incidence of monilia in his crop is nowadays less than 5%, producing a positive effect on his bottom line. I am hopeful that next time I visit Omar, he will have finished grafting his unproductive cocoa trees, his timber trees will be a few feet higher and hopefully his neighbors have also made similar progress because Omar’s success is evident.
Nutrition had a very nice and comprehensive editorial on the essence of chocolate. It mentions the recently discovered profitable effects of chocolate on the human physiology often mentioned on this blog.
In short:
Reduction of blood pressure by 6 grams of dark chocolate per day. Probably due to the flavonol epicatechin
Reduction of platelet and endothelial cell activation
Reduction of inflammatory mediators
It can also inhibit oral caries
It can cross the blood brain barrier and increase cerebral blood flow in humans
Rightly the authors question the effects of chocolate on mood and it’s possible addictive potential. As written before on this blog, chocolate is not an antidepressant and chocolate craving is a difficult concept consisting of different features.
The authors explain t...
Nutrition had a very nice and comprehensive editorial on the essence of chocolate. It mentions the recently discovered profitable effects of chocolate on the human physiology often mentioned on this blog.
In short:
Reduction of blood pressure by 6 grams of dark chocolate per day. Probably due to the flavonol epicatechin
Reduction of platelet and endothelial cell activation
Reduction of inflammatory mediators
It can also inhibit oral caries
It can cross the blood brain barrier and increase cerebral blood flow in humans
Rightly the authors question the effects of chocolate on mood and it’s possible addictive potential. As written before on this blog, chocolate is not an antidepressant and chocolate craving is a difficult concept consisting of different features.
The authors explain t...
Entry: Bill Guyton and Robert Peck, World Cocoa Foundation
Medellín, Colombia is situated in a valley about 45 minutes from the airport. This is the headquarters for World Cocoa Foundation member Compañía Nacional de Chocolates (CNCH) who manufactures chocolate products from bean to bar. We had time to tour the CNCH plant and learn more about their sourcing practices and outreach to cocoa farmers. Below is a photo taken from the entrance of the CNCH factory.
Robert Peck and Bill Guyton join Juan Fernando Valenzuela and colleague Ricardo for a tour of the CNCH facilities.
Our thanks to Sol Beatríz Arango, Enrique Escobar and Juan Fernando Valenzuela of CNCH for hosting us in Medellín.
As we leave Colombia, we are encouraged by what we have seen. An estimated 125,000 hectares of cocoa is currently under cultivation, grown by nearly 40,000 smallholder farmers. Although production is still around 38,000 metric tons, new planting areas will come into production over the coming years. It is estimated that by 2014, Colombia will become a net exporter of cocoa.
The efforts of both public and private sector cocoa initiatives have proven successful. The U.S. Agency for International Development (USAID) and several other development agencies have supported important programs over the past few years.
We at the World Cocoa Foundation look forward to continued collaboration with our member companies in Colombia, as well as the Cocoa National Council, a public-private sector organization, based in Bogota.
Entry: Bill Guyton and Robert Peck, World Cocoa Foundation
Manizales is a beautiful part of Colombia, best known for its coffee production. As you can see in the photo below, the rolling hillsides are covered with coffee trees which are the economic backbone for this region of Colombia.
Manizales, land of coffee
Although cocoa is not typically grown in Manizales, Robert and I are here to meet with World Cocoa Foundation member Casa Luker who support cocoa and agoforestry research at their farm less than an hour outside the city.
We are joined by Casa Luker representatives, Juan Carlos Arroyave and Alberto Agudelo for the journey. At the farm, we are introduced to Farm Manager Pedro Castellanos who brings us on a tour of the cocoa nurseries and training facilities.
Farm Manager Pedro Castellanos shows us grafted cocoa seedling.
The Casa Luker research facility has dormitories and classrooms dedicated to farmer training. Three-day training courses include topics on global marketing of cocoa, quality, planting and harvesting techniques, disease and pest management, and agroforestry.
Farmers can purchase grafted cocoa seedlings at the research facility. Most of the planting materials are Trinitario varieties, although CCN-51 is also available.
The greatest challenge to cocoa farmers in Colombia is containing diseases such as
Frosty Pod Rot (monilia). During the training at Casa Luker farm, participants learn about early detection of the disease and how to contain the spread. Certain varieties of cocoa are more resistant or tolerant to the disease.
After touring the training facilities, we walked through some of the agroforestry trials. Here the researchers are determining how best to plant cocoa with other trees such as bananas, plantains, papaya, passion fruit, coffee and timber species. Most of the spacing was roughly 3 x 3 meters for the fruit trees, while timber was intercropped at 4 x 16 meters.
From a smallholder farmer perspective, this type of system makes sense, since bananas can be harvested in the first couple of years, while cocoa matures and is ready to harvest starting in 4-5 years. Timber species take longer to mature, of course.
Cocoa grows well with many different types of tree species, including papaya.
We also saw some intensive cocoa plots with trees kept at 3-4 meters high which were producing well and without disease problems.
After completing the tour at the research farm, Robert and I were invited to visit the surrounding coffee farms and to see a local coffee processing facility.
We greatly appreciate the time spent with our colleagues at Casa Luker.
Recovering alcoholics and addicts have a long history of drinking large amounts of coffee, tea and cola soft drinks.
Coffee, tea, cola drinks, chocolate and many foods contain caffeine. A new range of drinks on the market is a variety of high content caffeine drinks such as Red Bull.
Caffeine, a stimulant, is the most widely consumed drug. Caffeine has occasionally been considered a drug of abuse and has the potential for people to become addicted.
Signs of caffeine addiction are;
Coffee simply stimulates the central nervous system, increases stress hormones in the blood streams, thus making a person feel unnaturally alert. Consequently increased alert state tends to subdue your body’s natural instincts and prevent it from relaxing. This causes undue stress and leads to various kinds...
Recovering alcoholics and addicts have a long history of drinking large amounts of coffee, tea and cola soft drinks.
Coffee, tea, cola drinks, chocolate and many foods contain caffeine. A new range of drinks on the market is a variety of high content caffeine drinks such as Red Bull.
Caffeine, a stimulant, is the most widely consumed drug. Caffeine has occasionally been considered a drug of abuse and has the potential for people to become addicted.
Signs of caffeine addiction are;
Coffee simply stimulates the central nervous system, increases stress hormones in the blood streams, thus making a person feel unnaturally alert. Consequently increased alert state tends to subdue your body’s natural instincts and prevent it from relaxing. This causes undue stress and leads to various kinds...
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Penn State Endowment Program faculty and students
I have the opportunity to visit Penn State University each fall, for an annual review of the program. It is encouraging to learn about the accomplishments of the program and the collaborations the PSU team has formed with researchers in cocoa-producing countries. Some of these achievements include the development of vegetative propagation methods that can be used in combination to achieve a large and rapid multiplication of cacao plants from single elite trees including in vitro tissue culture. The PSU program is also helping to identify ways of reducing disease losses on cocoa farms, as highlighted in one of our past blogs by PSU doctoral graduate, Rachel Melnick.
The World Cocoa Foundation is pleased to support the PSU cocoa research endowment with our member companies and thank Dr. Mark Guiltinan, Siela Maximova, the faculty and students who have made the program a success.
Omar is definitely a success story. Due to the amount of work he needs to invest into his farm, he has had to hire neighbors to help him with the agricultural practices that he has adopted. He is fortunate to have enough supply of water on his farm, which in the last year had been critical for his crops to thrive despite the dry season. The incidence of monilia in his crop is nowadays less than 5%, producing a positive effect on his bottom line. I am hopeful that next time I visit Omar, he will have finished grafting his unproductive cocoa trees, his timber trees will be a few feet higher and hopefully his neighbors have also made similar progress because Omar’s success is evident.
Farm Manager Pedro Castellanos shows us grafted cocoa seedling.
The Casa Luker research facility has dormitories and classrooms dedicated to farmer training. Three-day training courses include topics on global marketing of cocoa, quality, planting and harvesting techniques, disease and pest management, and agroforestry.
Farmers can purchase grafted cocoa seedlings at the research facility. Most of the planting materials are Trinitario varieties, although CCN-51 is also available.
The greatest challenge to cocoa farmers in Colombia is containing diseases such as Frosty Pod Rot (monilia). During the training at Casa Luker farm, participants learn about early detection of the disease and how to contain the spread. Certain varieties of cocoa are more resistant or tolerant to the disease.
After touring the training facilities, we walked through some of the agroforestry trials. Here the researchers are determining how best to plant cocoa with other trees such as bananas, plantains, papaya, passion fruit, coffee and timber species. Most of the spacing was roughly 3 x 3 meters for the fruit trees, while timber was intercropped at 4 x 16 meters.
From a smallholder farmer perspective, this type of system makes sense, since bananas can be harvested in the first couple of years, while cocoa matures and is ready to harvest starting in 4-5 years. Timber species take longer to mature, of course.