Progress 07/01/08 to 06/30/11
OUTPUTS: Four patents have been issued and two more patent applications have been filed. 42 biotech/biosafety regulators from developing and middle income countries received intense training on best practices for cost-effectively regulating genetically-modified crops and foods. AgBioForum.org, an online/electronic journal, has published more than 400 peer-reviewed articles since its beginning in 1998 and has a subscriber list of 4,810. The site reached more than 200,000 unique site visitors in 2010. PARTICIPANTS: Research scientist from the University of Illinois, University of Missouri and the Danforth Plant Sciences Center conducted the research reported in this Final Report. The researchers had collaborations with USDA-ARS, USDA-APHIS, FDA and EPA scientists. TARGET AUDIENCES: The goal of this grant was to produce corn and soybean products with commercialization potential. Agricultural producers and agribusinesses that develop, handle and market plant biotechnology products are the primary audiences. Additional audiences are consumers, domestic and international plant biotechnology regulators and members of the general public with interest in plant biotechnology. PROJECT MODIFICATIONS: Not relevant to this project.
A Missouri project identified soybean germplasm with bioactive lunasin levels 3X commonly found in widely adopted varieties. Lunasin affinity for hypoacetylated chromatin suggests a role in chromatin modification and in tumor suppressor in carcenogenesis. The goal of a soybean that yields "commercial nutraceutical levels" of lunasin that might provide cancer-preventive and cardiovascular benefits was attained. Danforth Center and University of Illinois scientists had goals (a) increasing the proline content of soybean seed to support the production of high proline fish tissues (i.e., skin and flesh) and (b) enhancing seed carotenes, especially astaxthanin, to provide pink flesh. Grant funds were matched by a European company with commercial interest in the new lines developed. The transgenic soybeans produced record amounts of beta-carotene in soybean and is first to improve proline content in plants as a near essential amino acid providing a foundation for designer soybean optimized as fish feed. A Missouri project developed technologies that enable designing new forms of soybean resistance to Asian soybean rust. The research team identified 20 peptides from among 250 that inhibit rust fungal spore germination/development and invented unique scaffolds to display peptides that inhibit rust pathogens. Four patents have been issued. Illinois scientists discovered the mechanisms soybean plants use to partition nutrient resources between protein and oil content at the molecular level. This information was used to increase the protein or oil content by genetic engineering approaches and/or improve the total yield. The methods employed were microarrays and Next Generation sequencing transcriptomics technologies to define the genetic program that begins in the stages of early seed development within a few days after flowering and fertilization. Project provided essential information to identify the gene networks for setting protein and oil composition. To understand gene expression networks leading to functional properties of the soybean seed, the researchers completed a detailed examination of soybean seed development during the stages of major accumulation of oils, proteins, and starches, as well as the desiccating and mature stages, using microarrays consisting of up to 27,000 soybean cDNAs. Gene expression data on seed development was published (Jones et.al., 2010) and entered at the National Center for Biotechnology Information at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgiacc=GSE18620. These data allow other researchers to access data sets on the complement of genes expressed in seed development. This grant partially supported the electronic journal, AgBioForum.org. ABF is the most widely read socio-economic journal on agricultural biotechnology. The journal had more than 200,000 unique reader visits in 2010. Two immersion courses on biotechnology regulation for developing country ministry-level regulators were offered (2009 and 2011). The $20,000 grant funding was matched with $290,000 in external funds and $100,000 of in-kind support. Forty-two regulators received in-depth training that is not available anywhere else.
- Fang, Z.D., Schoelz, J.E., Stacey, G., Schmidt, F.J. and English, J.T. 2008. Selection of phage-displayed peptides that inhibit soybean rust. American Phytopathological Society, Annual Conference. Minneapolis, MN. Phytopathology 98:S52.
- Fang, Z.D., Marois, J.J., Stacey, G., Schoelz J.E., English, J.T. and Schmidt, F.J. 2010. Combinatorially selected peptides for protection of soybean against Phakopsora pachyrhizi. Phytopathology: 100:1111-1117.
- Jones, S.I., Gonzalez, D.O. and Vodkin, L.O. 2009. Transcript profiling of soybean seed development from fertilization to maturity. Plant and Animal Genome XVI, p657.
- Jones, S.I., Gonzalez, D.O. and Vodkin, L.O. 2010. Flux of transcript patterns during soybean seed development. BMC Genomics 11:136.
- de Mejia, E.G. and V.P. Dia. 20910. Chemistry and biological properties of soybean peptides and proteins. In: Chemistry, Texture and Flavor of Soy. ACS Series, K. Cadwallader and S. Chang, Eds; ACS Symposium Series 1059, Chapter 9; American Chemical Society: Philadelphia, USA, 2010.
- Smehilova, M., Galuszka, P., Bilyeu, K.D., Jaworek, P., Kowalska, M., Sebela, M., Sedlarova, M., English, J.T. and Freobrt, I. 2009. Subcellular localization and biochemical comparison of cytosolic and secreted cytokinin dehydrogenase enzymes from maize. J. Exp. Bot. 60:2701-2712.
- Tuteja, J.H., Zabala, G., Varala, K., Hudson, M. and Vodkin, LO. 2009. Endogenous, tissue-specific short interfering RNAs silence the chalcone synthase gene family in Glycine max seed coats. Plant Cell 21: 3063-3077.
- Valliyodan, B. and Nguyen, H.T. 2011. Biological mechanisms that influence soy protein concentration and composition. In: Designing Soybeans for 21st Century Markets. R. Wilson, Ed; Chapter 8; American Oil Chemists Society, AOCS, Illinois, USA, (In Press).
- Widholm, J.M., Finer, J.J, Vodkin, L.O., Trick, H.N., LaFayette, P., Li, J. and Parrott, W. 2010. Transgenic plants in breeding and crop production - soybean transformation. In: F. Kempken and C. Jung (eds.) Genetic Modification of Plants, Biotechnology in Agriculture and Forestry 64, Springer-Verlag:Berlin Heidelberg pp. 473-498.
- Zernova, O., Zhong, W., Zhang, X.-H. and Widholm, J.M. 2008. Tissue culture specificity of the tobacco ASA2 promoter driving hpt as a selectable marker for soybean transformation selection. Plant Cell Reports 27:1705-1711.
- Zernova, O.V., Lygin, A.V., Widholm, J.M. and Lozovaya, V.V. 2009. Modification of isoflavones in soybean seeds via expression of multiple phenolic biosynthetic genes. Plant Physiol. Biochem. 47:769-777.
Progress 07/01/09 to 06/30/10
OUTPUTS: Projects supported this year focused on creating marketable products from corn and soybean. The projects ranged from increasing oil content in soybean to influencing the agricultural biotechnology regulatory process in countries that are deemed potential importers of corn and soybeans with biotechnology traits, commonly referred to as genetically-modified (GM) traits. PARTICIPANTS: From the University of Illinois: Drs. Lila Vodkin, Jack Widholm and Elvira de Mejia. From the University of Missouri: Drs. Henry Nguyen, David Sleper, Grover Shannon, James English, Frank Schmidt and Nicholas Kalaitzandonakes. From the Danforth Plant Sciences Center: Dr. Eliot Herman. TARGET AUDIENCES: U.S. consumers of food and fuel, consumers in developing countries that may become new purchasers of U.S. produced GM corn and soybean, and U.S. seed companies and international regulators of agricultural biotechnology crop imports are the targets of the research being conducted under this grant. PROJECT MODIFICATIONS: The decision to use a small portion of the grant funds to better understand the market barriers being placed on GM corn and soybean was a project modification of significance. This investment was matched six to one by external funds.
A. Illinois scientists are using functional genomics tools to create soybean with higher oil content. The research team discovered genes expression levels for cell growth and maintenance decreased as the cotyledons approached the mature, dry stage. Storage proteins genes had highest expression levels at the stage of highest fresh weight. Genes encoding many transcription factors and DNA binding proteins showed higher expression levels in the desiccating and dry seeds than in most of the green stages. Approximately 20 million sequences from each of six development stages were obtained: whole seeds at 4 days after flowering (DAF), whole seeds at 12-14 DAF; whole seeds at 5-6 mg fresh weight; seed cotyledons at 100-200 mg fresh weight, seed cotyledons at 400-500 mg fresh weight, and the mature dry seed cotyledon. From the high-throughput sequence data, the researchers were able to observe many changes in the transcript patterns over the six stages examined, i.e., some genes produce over 10,000 transcript copies while others occur only a few times. B. Illinois and Missouri investigators are developing new soybean varieties with significantly elevated levels of Lunasin, a peptide with potential in cancer prevention and protection against cardiovascular disease. The researchers have two years of data. C. A Missouri team is commercializing a technology to deliver defense peptides that protect soybean from Asian rust. In 2009, they attached defense peptides to a CKX (cytokinin oxidase/dehydrogenase from maize) scaffold and tested the displayed peptides for inhibition of rust when mixed with pathogen spores that are inoculated to soybean leaves. They produced significant reductions of infection with delays in symptom development. The results are published in the October 2010 issue of Phytopathology. The products of this research are two issued patents and a start-up company. D. An aquafeed project at the Danforth Plant Sciences Center has two components: 1. production of astaxthanin for the color of fish flesh and 2. increased proline content of soybean seeds to produce high proline fish skin and flesh tissue. Proline is a near essential amino acid in fish; there have been no prior attempts to improve proline content in seeds. The genes for each of these traits have been synthesized and placed into vectors and used to transform soybeans. For astaxanthin a high beta-carotene seed has been produced with RECORD levels of biotech produced b-carotene. B-carotene is the direct precursor of astaxanthin and can be produced by either a single or two-gene strategy; both strategies are being employed. To raise proline levels a proline synthesis gene with mutated allosteric feed back inhibitor has been placed in soybean and those transgenics are in selection. To confirm the mutated gene is expressed and effective, it has been placed in Arabidopsis; results show the gene is functional in vivo.
- Jones, S.I., Gonzalez, D.O. and Vodkin, L.O. 2010. Flux of transcript patterns during soybean seed development. BMC Genomics 11:136. (15 pages, online journal).
- Fang, Z.D., Marois, J.J., Stacey, G., Schoelz, J.E., English, J.T. and Schmidt, F.J. 2010. Combinatorially selected peptides for protection of soybean against Phakopsora pachyrhizi. Phytopathology 100:1111-1117.
Progress 07/01/08 to 06/30/09
OUTPUTS: Projects supported this year focused on creating marketable products from corn and soybean. The projects ranged from increasing oil content in soybean to influencing the agricultural biotechnology regulatory process in countries that are deemed potential importers of corn and soybeans with biotechnology traits, commonly referred to as genetically-modified (GM) traits. PARTICIPANTS: From the University of Illinois: Drs. Lila Vodkin, Jack Widholm and Elvira de Mejia. From the University of Missouri: Drs. Henry Nguyen, David Sleper, Grover Shannon, James English, Frank Schmidt and Nicholas Kalaitzandonakes. TARGET AUDIENCES: U.S. consumers of food and fuel, consumers in developing countries that may become new purchasers of U.S. produced GM corn and soybean, U.S. seed companies and international regulators of agricultural biotechnology crop imports are the targets of the research being conducted under this grant. PROJECT MODIFICATIONS: The decision to use a small portion of the grant funds to better understand the market barriers being placed on GM corn and soybean was a project modification of significance. This investment was matched 6 to 1 by external funds.
At the University of Illinois functional genomics tools are being employed to create soybean with higher oil content as oil is the more valuable constituent of soybean seed. Microarray and sequencing transcriptomics techniques are being used. Also, at Illinois and the University of Missouri investigators are developing new soybean varieties with significantly increased levels of Lunasin, a peptide that has potential in cancer prevention and protection against cardiovascular disease. Researchers in Missouri are commercializing a technology to deliver defense peptides to soybean that will protect them for Asian rust. The products of this research are nearing commercialization and will be high-yielding, high-value, disease resistant soybean varieties. In the area of regulatory studies, Missouri economists have assessed the barriers and opportunities in markets where GM corn and soybean are not yet accepted, but have high potential for acceptance. A significant success was a 10-day international immersion course on agricultural biotechnology regulation attended by 19 ministry-level regulators from 13 countries in Africa, Asia and Eastern Europe. The USDA-FAS and QUALISOY industry council were partners in the immersion course. The 19 participants are now capable of providing objective opinions and counsel as their governments address the critical issue of accepting GM crops into their markets.
- No publications reported this period