Progress 07/01/10 to 06/30/14
Outputs
Target Audience: General scientific community. Chocolate industry. Cacao scientists in developing countries. USAID and other development agencies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Five graduate students and 8 undergraduate student gained experience in plant molecular biology research as a result of this project period. Faculty and staff gained professional development in leadership and organizational skills in running the project and in several formal training sessions. How have the results been disseminated to communities of interest? Peer reviewed publications. Speaking and poster sessions and US and international meetings. Outreach to broader community: As part of this endowed program, representatives of key companies attend meetings twice yearly to review recent progress and discuss issues with Dr. Guiltinan and University administrators related to the industry needs and outlooks. Feedback from industry is essential in connecting our research to stakeholders needs, and in networking between industry and academic labs. Industry partners will be updated twice a year on the progress of the proposed project via this forum. The international cacao research community consists of many groups geographically dispersed with many different collaborative initiatives. It is important for the impact of our research that other cacao scientists are aware of and contribute to our projects where appropriate. To facilitate this, this project will be highly interactive with all international cacao research organizations and the National Plant Genomics Initiative community through the active participation of the PI and co-PIs and other project participants with the following organizations. International Group for Genetic Improvement of Cocoa (INGENIC) INGENIC is a worldwide group of approx. 300 cacao breeders and geneticists. It promotes exchange of information and collaboration in the area of cacao genetics and breeding worldwide. The PI (Guiltinan) is a member of the INGENIC Executive committee and chair of the Study group on Cacao Molecular Genetics. Through the INGENIC website, email list-serves, and our biannual meetings, the INGENIC research community will be updated about the progress of this project regularly. The World Cocoa Foundation (WCF) WCF is a 501 (c) 3 organization dedicated to improving the standard of living of cacao farmers around the world by providing training on low-cost methods to produce quality cocoa in a sustainable, environmentally friendly manner. It is supported by donations from private and corporate sponsors. This organization is particularly active in Africa and Asia in farmer educational and other outreach activities. We anticipate that the WCF will make important contributions towards the training component of this proposal through its extensive networks throughout the developing world and high visibility in the international development community. For example, WCF administers a Borlaug Fellows program and has funded scientific exchange visits to the PIs lab by several West African Scientists. The PI will communicate with the WCF via attendance at WCF Partnership meetings yearly. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
To evaluate the efficacy of several strategies for anti-pathogen resistance in cacao. Results: Developed and testing several different transgenes targeted at pathogen resistance. Developed transient expression system that allows for fast preliminary screening of selected candidate gene and their products against fungal and oomycete infection in cacao green leaf tissue. Several potential candidate genes were evaluated and their over-expression has resulted in significant increase of resistance of transgenic cacao plants against Phytophthora palmivora, a major pathogen of cacao (manuscript in review by Plant Biotechnology Journal). Scientific exchange visits concluded with scientists from Cameroon, Panama, Mexico, Ghana and Ivory Coast. Objectives: increasing the success of cacao somatic embryogenesis of valuable cacao genotypes by investigating the basic mechanism of somatic embryo regeneration and creating low-cost, high throughput bioreactors. Results: Whole genome expression microarray was manufactured and used to hybridize RNA from 24 samples representing different developmental stages of zygotic and somatic embryos. Global gene expression profiles and the profiles of key seed-specific and embryo development transcription factors, seed storage proteins, fatty acid biosynthesis, and flavonoid genes are in the process of evaluation. New bioreactor design implemented and in testing with cacao. Functional Genomic Tool Development for Cacao: We have developed functional genomics tools for cacao including a stable transformation system, Agrobacterium infiltration transient leaf expression assay, whole genome microarray and predicted proteome peptide database for mass spectrometry based protein identification (unpublished data). We have used transgenic Arabidopsis, tobacco and tomato to study the function of different cacao genes including genes involved in the defense and flavonoid pathways. Perhaps the most powerful method we have adapted for cacao functional studies is a transient expression system that was recently developed with funding from the NSF-BREAD program.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Liu, Y., Shi, Z., Maximova, S., Payne, M. & Guiltinan, M. J. Proanthocyanidin synthesis in theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase. BMC Plant Biology 13, 202, doi:10.1186/1471-2229-13-202 (2013).
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Shi, Z., Zhang, Y., Maximova, S. & Guiltinan, M. J. TcNPR3 from Theobroma cacao functions as a repressor of the pathogen defense response. BMC Plant Biology 13, 204, doi:DOI: 10.1186/1471-2229-13-204 (2013).
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Yi, L., Zi, S., S, M., Payne, M. & Guiltinan, M. J. Proanthocyanidin synthesis in theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase. BMC Plant Biology 13, 202, doi:10.1186/1471-2229-13-202 (2013).
|
Progress 10/01/12 to 09/30/13
Outputs
Target Audience: General scientific community. Chocolate industry. Cacao scientists in developing countries. USAID and other development agencies. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Educational Impact: Three graduate students and three undergraduates were involved in this research this year. Three technicians and one research associate were also involved. During this period we hosted two international scientists who each completed research projects. The ACRI endowment supported these visits by funding supplies and technical support. The Penn State College of Agricultural Sciences also provided some matching support for these visits. How have the results been disseminated to communities of interest? Through publications, and presentations at numerous scientific meetings and workshops. Through display at local science museum. Through our website. What do you plan to do during the next reporting period to accomplish the goals? Continue with the same projects. Writing proposals to expand funding base. Working closely with industry partners to establish new projects and expand our impact. We are working with the Smithsonian Tropical Research Institute to try to establish a long-term field site for colalborative research on cacao farming sustainability.
Impacts
What was accomplished under these goals?
Project I Molecular Biology of Oil Biosynthesis in Theobroma cacao This year we have made several advances in understanding which genes control the key steps in cocoa butter biosynthesis. In particular we have identified some of the key genes involved in controlling the formation of double bonds which determine to a large degree the physical properties of cocoa butter. Project II Sequencing the Genome of Cacao We continue to explore the cacao genome mining it for the genes most important to disease resistance and seed quality traits. Project III Improvement of Cacao Somatic Embryogenesis With leveraged funding from the National Science Foundation, we continue to develop the tissue culture systems for cacao to make it more efficient in genotype independent. This will have impact in the near future as commercial cacao tissue culture labs are rolled out. Our technology has been or is being adopted worldwide. One significant recent finding is that the gene Lec2 is critical for the induction of embryogenesis in cacao. We have determined the expression patterns of thousands of genes that seemed to be involved in the SE process an din normal zygotic embryogenesis. Project IV Molecular Biology of Defense Response in Cacao Using a combination of approaches we are investigating the mechanisms by which cacao responds to pathogen infection and switches on its defense response. We have used several genomics approaches to identify key genes involved in defense pathway in cacao. We have developed microarrays that allow us to measure the expression of tens of thousand cacao genes in a single experiment. We have also made use of the model plant Arabidopsis to isolate and study key cacao genes involved in regulating the defense response. We have leveraged funding from the National Science Foundation, in collaboration with the Bill and Melinda Gates Foundation, to expand these studies into testing of novel disease resistance approaches.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Bailey, B.A., Crozier, J., Sicher, R.C., Strem, M.D., Melnick, R., Carazzolle, M.F., Costa, G.G.L., Pereira, G.A.G., Zhang, D., Maximova, S., Guiltinan, M., and Meinhardt, L. (2013). Dynamic changes in pod and fungal physiology associated with the shift from biotrophy to necrotrophy during the infection of Theobroma cacao by Moniliophthora roreri. Physiological and Molecular Plant Pathology 81, 84-96.
|
Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: This is a summary of an ongoing research project on cacao molecular biology. The main work during this period has been the continued focus on elucidation of cacao disease resistance mechanisms and to begin to explore genes for important quality traits. We have developed several strategies for rapid functional testing of cacao gene function. Using these translational biology approaches, we have begun to characterize resistance genes, as well as genes in the lipid and flavonoid biosynthetic pathways. Our main findings include isolation and characterization of the key plant immune system regulatory genes NPR1 and NPR3, as well as the major structural genes for all of the key steps in the lipid and flavonoid pathways. We have provided training to four graduate students, two postdoctoral scholars, many undergraduates and several visiting scientists from Mexico, Panama and Cameroon. PARTICIPANTS: We have provided training to four graduate students, two postdoctoral scholars, many undergraduates and several visiting scientists from Mexico, Panama and Cameroon. TARGET AUDIENCES: Scientists in academic and industry labs concerned with cacao genetics and breeding. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
This work has contributed to advancing the understanding of disease resistance mechanisms in cacao. We are developing collaborations to extend our work to screen large collections of germplasm for variation in gene expression for the genes described above. Ultimately this will lead to molecular markers for accelerating the pace of cacao breeding for disease resistance and quality traits. We will impact the development of research capacity in developing countries through continued hosting of visiting and collaborating scientists.
Publications
- Shi, Z., S. Maximova, Y. Liu, J. Verica, and M. J. Guiltinan. 2012. The Salicylic Acid Receptor NPR3 Is a Negative Regulator of the Transcriptional Defense Response during Early Flower Development in Arabidopsis. Molecular Plant http://dx.doi.org/10.1093/mp/sss091.
- Mejia, L., M. Guiltinan, Z. Shi, L. Landherr, and S. Maximova. 2012. Expression of Designed Antimicrobial Peptides in Theobroma cacao L. Trees Reduces Leaf Necrosis Caused by Phytophthora spp. In: Small Wonders: Peptides for Disease Control. American Chemical Society 1095:379-395.
- Costa, G. G. L., O. G. Cabrera, R. A. Tiburcio, F. J. Medrano, M. F. Carazzolle, D. P. T. Thomazella, S. C. Schuster, J. E. Carlson, M. J. Guiltinan, B. A. Bailey, et. al. 2012. The mitochondrial genome of Moniliophthora roreri, the frosty pod rot pathogen of cacao. Fungal Biology 116(5):551-562.
|
Progress 10/01/10 to 09/30/11
Outputs
OUTPUTS: This is a summary of an ongoing research project on cacao molecular biology. The main work during this period has been the continued focus on elucidation of cacao disease resistance mechanisms and involvement in the cacao genome project. The result of this work is the completed genome sequence and its initial description including the discovery of over 28,000 genes and many genes implicated in high value traits. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
This work has contributed to advancing the understanding of disease resistance mechanisms in cacao. The immediate impact of this work will be on helping accelerate and define new research directions.
Publications
- Argout, X., J. Salse, J. Aury, M. Guiltinan, G. Droc, J. Gouzy, M. Allegre, C. Chaparro, T. Legavre, S. Maximova, M. Abrouk, F. Murat, O. Fouet, J. Poulain, M. Ruiz, Y. Roguet, M. Rodier-Goud, J. Barbosa-Neto, F. Sabot, D. Kudrna, J. Ammiraju, S. Schuster, J. Carlson, E. Sallet, T. Schiex, A. Dievart, M. Kramer, L. Gelley, Z. Shi, A. Berard, C. Viot, M. Boccara, A. Risterucci, V. Guignon, X. Sabau, M. Axtell, Z. Ma, Y. Zhang, S. Brown, M. Bourge, W. Golser, X. Song, D. Clement, R. Rivallan, M. Tahi, J. Akaza, B. Pitollat, K. Gramacho, A. Dhont, D. Brunel, D. Infante, I. Kebe, P. Costet, R. Wing, W. McCombie, E. Guiderdoni, F. Quetier, O. Panaud, P. Wincker, S. Sidibe-Bocs, and C. Lanaud. 2010. The genome of Theobroma cacao. Nature Genetics. 26 December 2010. http://dx.doi.org/10.1038/ng.736.
- Aikpokpodion, P., M. Kolesnikova-Allen, V. Adetimirin, M. Guiltinan, A. Eskes, J. Motamayor, and R. Schnell. 2010. Population structure and molecular characterization of Nigerian field genebank collections of cacao, Theobroma cacao L. Silvae Genetica 59:273-289.
- Shi, Z., S. Maximova, Y. Lui, J. Verica, and M. Guiltinan. 2010. Functional Analysis of the Theobroma cacao NPR1 Gene in Arabidopsis. BMC Plant Biology 10-248.
- Micheli, F., M. Guiltinan, K. Gramacho, M. Wilkinson, A. Figueira, J. Cascardo, S. Maximova, and C. Lanaud. 2010. Functional Genomics of Cacao. In Advances in Botanical Research. J.-C. Kader and M. Delseny, eds (Burlington: Academic Press). pp. 119-177.
|
Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: This is a summary of an on-going research project on cacao molecular biology. Our main outputs during this period involved publications from graduate students and finishing of one Ph.D. student. Four graduate students and two undergraduates were involved in this research. Two technicians and one research associate were also involved. We hosted two international scientists who each completed research projects while in our lab for periods of up to one year. These were supported with various funds including a Fulbright Scholarship and funds from the home country research institutes. PARTICIPANTS: Department of Horticulture: Kenzakoski, Marissa; Maximova, Siela; Mejia, Franco; Melnick, Luis; Pishak, Sharon; Shi, Zi; Wahid, Nur; Sabrina, N.; Young, Ann; Zhang, Yufan. Collaborators USDA Beltsville, Smithsonian Institute, University of Maryland, Virginia Biotechnology Institute. TARGET AUDIENCES: Cacao genetics research community Plant biology researchers Chocolate manufacturing companies Federal granting agencies (NSF). PROJECT MODIFICATIONS: During the past year we received a grant from the NSF that has resulted in a change to include collaborative research aimed at testing possible approaches to fungal resistance.
Impacts
This work has contributed to advancing the understanding of disease resistance mechanisms in cacao. The immediate impact of this work will be helping to accelerate and define new research directions.
Publications
- Cakirer, M., G. Ziegler, and M. Guiltinan. 2010. Seed Color as an Indicator of Flavanol Content in Theobroma cacao L. In Chocolate Consumption and Health, Hauppauge, NY. Nova Publishers. pp. 257-270.
- Marelli, J. P., S .N. Maximova, K. P. Gramacho, S. Kang, and M. J. Guiltinan. 2009. Infection biology of Moniliophthora perniciosa on Theobroma cacao and alternate solanaceous hosts. Tropical Plant Biology 2:149.
- Tiburcio, R., G. Costa, M. Carazzolle, J. Mondego, S. Schuster, J. Carlson, M. Guiltinan, B. Bailey, P. Mieckowski, L. Meinhardt, and G. Pereira. 2009. 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. Jan. Journal of Molecular Evolution 70(1):85-97.
|
Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: NPR1 control of defense gene activation: NPR1 is a key protein transcriptional regulator of defense response. It has been extensively characterized in Arabidopsis, a model plant for molecular-genetic research. In Arabidopsis, NPR1 resides in the cytoplasm, but upon perception of pathogen infection, NPR1 quickly moves into the nucleus where it interacts with a number of different proteins that together activate about 120 genes involved in plant defense response. As a result of the various actions of these proteins (defense response proteins) a defense response is mounted, leading in some cases to an inhibition or reduction in the ability of the pathogen to invade. We have isolated and begun to characterize the NPR1 gene of cacao. This gene shows a very high degree of similarity to the Arabidopsis gene. Using a technique called transgenic complementation, we have moved this gene into Arabidopsis plants that lack a native NPR1 gene, and have shown that the cacao gene functions similarly to the Arabidopsis gene. The flavonoids are a large family of compounds important in many functions during plant development including pest and disease resistance. This class of molecules includes the proanthocyanidins, which are well known for their health promoting activities in chocolate. We have isolated several of the key genes that participate in the synthesis of these compounds and are studying their expression. One interesting finding is that several of these genes are expressed early in leaf development but as leaves mature, the genes are shut off. Genome Sequencing of Cacao Pathogens: Determining the sequences of plant pathogens reveals clues to the mechanisms of pathogenesis and allows scientists to study the genetic diversity of different pathogen strains. In collaboration with USDA Beltsville (L. Meinhardt and B. Bailey) and UNICAMP, Brazil, (G. Pereira), along with collaborators at Penn State (S. Schuster and J. Carlson) we have sequenced and are now analyzing the entire genome sequences of two of the most devastating cacao plant pathogens, witches' broom and frosty pod. These two fungi are responsible for the majority of cacao crop losses throughout Central and South America. The completion of the genome sequences of these pathogens opens up new frontiers in understanding the mechanisms of pathogenesis and in developing strategies for increased resistance. PARTICIPANTS: Four graduate students and two undergraduates were involved in this research. Two graduate students received Ph.D. degrees recently. We hosted four international scientists who each completed research projects while in our lab for periods of up to one year. We collaborate extensively with Dr. Paul Backman in the Penn State Dept of Plant Pathology. TARGET AUDIENCES: The scientific community via publications and in the long run, cacao farmers in developing countries. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Four graduate students and two undergraduates were involved in this research. Two graduate students received Ph.D. degrees recently. We hosted four international scientists who each completed research projects while in our lab for periods of up to one year. These were supported with various funds including: Fulbright Scholarship, Borlaug International Fellowship, and funds from the home country research institutes.
Publications
- Marelli, J.P., S.N. Maximova, K.P. Gramacho, S. Kang, and M.J. Guiltinan. 2009. Tomato as a model system to study pathogenicity mechanisms of Moniliophthora perniciosa, the causal agent of witches' broom disease of cacao. Phytopathology. (Pending).
- Guiltinan, M.J., J. Verica, D. Zhang, and A. Figueira. 2008. Genomics of Theobroma cacao, the Food of the Gods. In Genomics of Tropical Crop Plants, P.A.M. Moore, R, ed. (New York: Springer). pp. 145-170.
- Maximova, S.N., A. Young, S. Pishak, and M. Guiltinan. 2008. Field performance of Theobroma cacao L. plants propagated via somatic embryogenesis. In Vitro Cell. Dev. Biol. - Plant 44:487-493.
- Maximova, S.N. and M.J. Guiltinan. 2008. Cocoa. In A Compendium of Transgenic Crop Plants, C. Kole and T. Hall, eds (Oxford, UK: Blackwell Publishing). pp. 85-98.
- Melnick, R., N. Zidack, B. Bailey, S. Maximova, M. Guiltinan, and P. Backman. 2008. Bacterial endophytes: Bacillus spp. from vegetable crops as potential biological control agents of black pod rot of cacao. Biological Control 46:46-56.
- Mondego, J., M. Carazzolle, G. Costa, E. Formighieri, L. Parizzi, C. Cotomacci, R. Vidal, L. Nascimento, R. Estrela, M. Guiltinan, B. Bailey, L. Meinhardt, J. Cascardo, and G. Pereira. 2008. A genome survey of Moniliophthora perniciosa gives new insights about cacao Witches Broom Disease. BMB Genomics 9:548.
- Swanson, J.D. and M.J. Guiltinan. 2008. Comparative flower development in Theobroma cacao based on temporal morphological indicators. International Journal of Plant Sciences 169, 1187-1199.
- Xavier-Argout, O.F., P. Wincker, K. Gramacho, T. Legavre, X. Sabau, A.M. Risterucci, C. Da Silva, J. Cascardo, M. Allegre, D. Kuhn, J. A. Verica, B. Courtois, R. G. Loor, B. Regis, O. Sounigo, M. Ducamp, M. J. Guiltinan, M. Ruiz, L. Alemanno, R. Machado, W. Phillips, R. Schnell, M. Gilmour, E. Rosenquist, D. Butler, S. Maximova, and C. Lanaud. 2008. Towards the understanding of the cocoa transcriptome: Production and analysis of an exhaustive dataset of ESTs of Theobroma cacao generated from various tissues and under various conditions. BMC Genomics 9:512.
|
|