Location: Commodity Utilization Research
2025 Annual Report
Objectives
As directed by ARS research priorities, this proposal is focused on four broad objectives.
Objective 1: Develop novel cottonseed oil products with traits to maintain and enhance market value.
Sub-Objective 1a. Develop G. hirsutum cotton germplasm with 40% oleic acid in its seed oil.
Sub-Objective 1b. Identify and characterize the additional genetic elements that contribute to the high oleic acid trait in GB713.
Sub-Objective 1c. Modify cyclopropane synthase genes to reduce the levels of CPFAs in cotton tissues.
Sub-Objective 1d. Modify and combine cotton and other plant genes to increase production of DHSA in lipids of roots, seeds, and other tissues.
Objective 2: Explore reported seed quality concerns to improve seed quality.
Objective 2a. Determine the magnitude and range of seed hull fracture resistance of the Gossypium species that produce usable cotton fiber.
Sub-objective 2b. Determine the relative importance of genetics, environment, and their interaction on the fracture resistance of cottonseed.
Sub-objective 2c. Develop a method to measure the propensity of cottonseed to be damaged during convening or ginning.
Sub-objective 2d. Study the rate of deterioration in the quality of whole and damaged cottonseed under different storage conditions.
Objective 3: Study the potential for using the whole seed and defatted protein meal of low-gossypol plant lines in food applications.
Sub-objective 3a. Develop acidic juices and drinks fortified with cottonseed protein.
Sub-objective 3b. Develop cottonseed-based butter and spread products.
Sub-objective 3c. Develop cottonseed protein-based food-grade films to improve food shelf life.
Objective 4: Develop new or modified processing methods to increase the value of processed products from cottonseed.
Objective 4a. Recover the tocopherol and sterol components of deodorization distillate and add these back to deodorized cottonseed oil to improve its stability.
Approach
Several analytical, chemical, physical, microbiological, and genetic techniques will be employed to achieve the project goals. Genetic manipulation, molecular biology, and classical breeding methods will be used to study the synthesis of the cyclopropyl fatty acids and to increase seed oil oleic acid levels. Gas chromatography will be used to determine oil fatty acid profiles, which are needed in several objectives. Various physical and chemical techniques will be employed at the laboratory level to study seed durability and hardness. Some developmental work will be needed to develop a technique that can be used to test for seed durability. Chemical and physical techniques will be used to formulate ingredients and food products from seed kernels and to isolate high protein fractions to use to generate film products. Some of these potential products will also be evaluated by sensory panels. The processing objective will utilize a number of chemical fractionation methods to either eliminate unwanted components or to extract potentially useful components from deodorizer distillate.
Progress Report
This is the final report for project 6054-41000-113-000D, Development of Novel Cottonseed Products and Processes, which was replaced by project 6054-30600-001-000D, Enhanced Cottonseed Profitability and Sustainability through Improved Genetics and Byproduct Formulation in July 2025.
The project focused on issues related to the processing of cottonseed components for food-based products, films, food packaging and other biobased products. The project objectives fall under National Program 306, Component 1 – Foods, Problem Statement 1A: Define, Measure, and Preserve/Enhance/Reduce Factors that Impact Quality and Marketability; Problem Statement 1.B: New bioactive ingredients and health-promoting foods; and Problem Statement 1.C: New and improved food processing and packaging technologies.
Cottonseed oil was once the ‘gold standard’ of frying oil. Over time, other vegetable oils gained market share due to higher stability at high frying temperatures and healthier chemical composition, due to higher levels of the ‘heart-healthy’ fatty acid called oleic acid. ARS researchers in New Orleans, Louisiana, together with ARS researchers at Mississippi State, Mississippi bred high oleic acid traits into seed oils of standard fiber-production upland cotton. Six lines with at least 50% oleic acid (300% more than standard cottonseed oil) were identified. The lines were screened for multiple generations and across multiple geographic locations to ensure stability of high oleic acid levels, normal seed fiber traits, and overall field performance. Three lines met the defined metrics, including 50-55% seed oleic acid. The description of these lines was recently published, and seeds were made available to private and commercial cotton growers. These resources will add value to the American agricultural economy and provide the U.S. cotton industry with a new asset to drive competitive advantage. However, it is not clear how many genes influence this trait. Recent research identified at least five novel gene versions that appear to be linked to the trait; statistical linkage testing for two was recently completed. Testing of the others is ongoing. The trait-linked genes can be used as ‘markers’ for future breeding programs. This will allow companies and private cotton growers to introduce the high oleic acid trait into commercial fiber producing varieties and easily track the linked genes, saving time and money.
Another novel aspect of cottonseed oil is the presence of an unusual three-carbon ring structure in fatty acids called CPFAs. Depending on the end usage, cotton varieties containing either more, or less, CPFAs than are found in standard cottonseed oil would benefit the cotton industry. Recent research identified cotton genes that enhance CPFA accumulation. Two enzymes were tested and found to increase CPFA levels in plants. Future creation of high CPFA cottonseed oil will economically benefit cotton growers and oil processors. Consumers will benefit from consumption of this oil, which will improve blood cholesterol levels.
On the other hand, low CPFA cottonseed oil would also be a useful commodity. Other experiments targeted the development of an efficient cotton genome editing system. The technology known as ‘CRISPR’ offers possibilities for the development of value-added non-transgenic cotton plants. However, this technology has not received much attention in cotton to date. Work conducted during this project made major progress towards a cotton leaf-based genome editing screening protocol. A two-week experiment using one cotton leaf can now better identify which editing strategies will be worthy of the 18-month commitment required for the creation of entire engineered cotton plants.
In support of Objective 2, the scientists conducted research to develop methods for detection of cottonseed damage that occurs during harvest and ginning. A low-cost, high-volume method would be very valuable to cotton producers by allowing for screening and removal of seed lots that would perform poorly in the field. The developed damage assessment method captures the solvent-extractable kernel components that leach from damaged seeds and measures the light absorption in each sample. Three levels of seed damage were compared. All three levels of damage could be detected, but the differences between minor and moderate damage could not always be distinguished. The source of seeds also influenced the outcomes. Due to staffing shortages, progress on this aspect of research slowed towards the end of this project. This was also true for the second component of Objective 2, which focused on development of standardized storage conditions that would optimize longevity and field performance of cottonseeds, including damaged seeds.
Cottonseed protein and cotton gin trash are underutilized byproducts generated from cotton production. In support of Objective 3, the ARS scientists collaborated with scientists at the Rochester Institute of Technology to investigate the usage of cottonseed protein as paper coating for food packaging applications. Various formulation properties were tested, including mechanical strength, and oxygen and water vapor barrier levels. Cottonseed protein-based coatings proved viable as a competitive paper coating. Cotton “gin trash”, another low-cost high volume processing byproduct, proved to be a valuable addition to cottonseed protein coating formulations. For a food packaging application that allows the food item in the package to “breathe”, mixed cottonseed protein/gin trash paper coatings showed moderately good performance. This work also pointed out possible future approaches that can further improve the performance of cottonseed protein-coated paper for food packaging.
Given the levels of plastic pollution in the environment and micro-plastics in water, research is urgently needed to replace synthetic plastics. In support of Objective 3, ARS scientists explored the properties of blended biodegradable polymers that contain washed cottonseed meal as possible agriculturally-based, biodegradable plastic alternatives called biocomposites. Plasticizer additives such as glycerol were also included in the tested formulations. Such films can be used as water-soluble food coatings and as pouches for detergents and agrochemicals. Additional research expanded this work to include other cotton byproduct feedstocks such as linters and seed oil. Four-component (resin,washed cottonseed meal, cottonseed oil, and glycerol) blends were optimized using a process called ‘melt compounding’. The results supported the use of these biocomposites in production of low-strength biodegradable nursery plant containers. Two Cotton Incorporated-funded projects (agreements #0000074382 and #0000074381) also supported the biocomposites research and expanded it to include the use of other cotton biomass byproducts in the production of particleboards.
Plant-based butters have steadily increased in consumer popularity. The ARS scientists made peanut butter-like food products from glandless cottonseed kernels. These cottonseed products were characterized by color, texture, and physical properties. Cottonseed producers and food manufacturers will benefit from marketing and sales of spreadable cottonseed butters. These products will compete with peanut butter, with less risk of allergic reactions in some consumers. Other recent research monitored and compared the shelf-life properties of food products containing crude and refined cottonseed oil under various storage conditions. This work suggested that inclusion of crude cottonseed oil improves the quality and shelf life of cottonseed oil-containing food products.
Improvement of glandless cottonseed protein isolate solubility was studied. Annual cottonseed processing generates enough protein to meet the needs of 500 million people, but the low solubility of cottonseed protein negatively affects its use in food and beverage applications. Protein was treated with a range of modifiers to improve its solubility. These experiments showed that positively charged food-grade surfactants, higher temperature and reduced protein particle size increased solubility, based on tests in three fruit juices and soda. This research provides useful guidelines to manufacturers for the development of protein-rich juices and drinks.
The researchers also studied cottonseed as a source of high-value bioactive materials for improving health and nutrition. Finding ways to reduce the occurrence of obesity and diabetes will reduce healthcare costs and improve quality of life. Cottonseed contains many ‘bioactive’ compounds. Ongoing research has studied the positive effects of cottonseed extracts on sugar metabolism in cells that regulate both insulin resistance and inflammatory reactions in fatty tissue.
The goal of Objective 4 was to recover valuable components from the deodorization distillate that is formed as a byproduct of cottonseed oil refining. This objective requires a source of deodorization distillate, which was to be acquired from a collaborator. Due to disruptions from the pandemic, staffing shortages, and the continued difficulty of obtaining commercial cottonseed deodorizer distillate, little progress was made towards these milestones.
Summarizing the accomplishments over the five year project cycle, substantial progress was made on the development of cottonseed lines with higher levels of heart-healthy seed oil oleic acid, exceeding the initial goals stated in the research plan. Progress towards creation of cottonseeds containing altered levels of cholesterol-modulating CPFA fatty acids was also achieved. Additional development of engineered lines will be a priority in the future.
Accomplishments
1. Public release of next-generation heart-healthy cottonseed oil genetic resources.. Decades ago, cottonseed oil was used as an ingredient in many food products, especially as a frying oil for potato chips and other popular snack foods. Over time, cottonseed oil lost market share to other vegetable oils with healthier and more stable chemical profiles. Working with ARS collaborators at Mississippi State, Mississippi, ARS researchers in New Orleans, Louisiana, completed the breeding and characterization of new cottonseed lines containing heart-healthy seed oil. These oils contain three times the level of the key fatty acid that is responsible for this trait, compared to normal cottonseed oil. The high oleic acid content also confers greater stability at high cooking temperatures and longer shelf-life during storage. This resource will create new food (and non-food) markets for cotton growers and seed processors and will enhance profitability of the U.S. cotton industry.
2. Roasting temperature impacts the quality and shelf life of cottonseed butter/spread products.. The chemical composition of cotton kernels is compatible with production of ‘cottonseed butter’, a high-protein spread that would contain much lower levels of allergens than peanut butter. Optimized cottonseed butter properties require proper roasting conditions. Roasting is one of the most important processes in food applications that rely on seeds or seed kernels. ARS researchers in New Orleans, Louisiana, observed that high roasting temperatures deepened the color and increased the firmness and spreadability, but decreased the oxidative stability of these cottonseed butter products. Knowledge derived from this work could help to optimize manufacturing processes and enhance preparation of better-quality cottonseed butter products for food companies with greater appeal for U.S. consumers.
3. Cottonseed byproducts and simple resins or polymers make useful biocomposite plastic substitutes.. Biocomposites are materials made by blending some type of matrix, usually a chemical resin, with renewable resources added as modifiers or fillers to match the specifications for various manufacturing applications, such as food and non-food packaging or baskets and pots for nursery plants. Using low-value, high volume cottonseed biomass products as fillers will lower biocomposite production costs and increase the profitability and resilience of cotton industries by creating new revenue streams for underused byproducts. The heat stability of biocomposite products is important for developing and managing strategies for processing, application, and recycling of these products. ARS researchers in New Orleans, Louisiana, collaborated with University of Massachusetts Lowell scientists to test the heat stability of 4-ingredient biocomposites containing cottonseed byproducts. Information derived from this work will reduce environmental plastic contamination by providing meaningful guidance for targeted stabilization and post-use recycling of these biocomposites.
4. Optimal conditions for increasing cottonseed protein solubility for fortified juices and drinks.. The cottonseed industry generates 10 million tons of protein annually, enough protein to meet the needs of half a billion people. However, the low solubility of cottonseed protein is a major factor affecting its use in most food and beverage applications. ARS researchers in New Orleans, Louisiana, developed optimal conditions for increasing the solubility of cottonseed protein, which is useful for developing ‘high protein’ formulations of drinks with acidic pH, such as fruit juices and sodas. This research opens new avenues for using cottonseed as a source of high-quality, plant-based protein in various food applications. This research made significant contributions to agricultural sustainability, food safety, and innovative product development by meeting the growing demand for sustainable, protein-rich alternatives in the domestic and global food supplies.
Review Publications
Mcguire, S.T., Shockey, J.M., Bates, P.D. 2024. The first intron and promoter of Arabidopsis DIACYLGLYCEROL ACYLTRANSFERASE 1 exert synergistic effects on pollen and embryo lipid accumulation. New Phytologist. 245(1): pg263-281. https://doi.org/10.1111/nph.20244.
He, Z., Cheng, H.N., Ford, C.V., Nam, S., Fortier, C., Santiago Cintron, M., Olanya, O.M., Uknalis, J. 2024. Four-ingredient blends of poly(lactic acid) with cottonseed oil and meal for biocomposite utilization. Macromol. 4(4): 708-722. https://doi.org/10.3390/macromol4040042.
Wang, Y., Zhu, Y., Guo, G., An, L., Fang, W., Tan, Y., Jiang, J., Bing, X., Song, Q., Zhou, Q., He, Z. 2024. A comprehensive risk assessment of microplastics in soil, water, and atmosphere: Implications for human health and environmental safety. Ecotoxicology and Environmental Safety. 285. Article 117154. https://doi.org/10.1016/j.ecoenv.2024.117154.
Pundir, A., Thakur, M.S., Prakash, S., Kumari, N., Sharma, N., Parameswari, E., He, Z., Nam, S., Thakur, M., Puri, S., Puranik, S., Kumar, S., Madhu, Kuma, M. 2024. Fungi as versatile biocatalytic tool for treatment of textile wastewater effluents. Critical Reviews in Environmental Science Technology. 36. Article 185. https://doi.org/10.1186/s12302-024-01007-3.
He, Z. 2025. Agricultural and environmental significance of soil organic matter and plant biomass: Insight from ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry. Pedosphere. 35(1). pg3-7. https://doi.org/10.1016/j.pedsph.2024.09.004.
Bates, P.D., Shockey, J.M. 2024. Towards rational control of seed oil composition: dissecting cellular organization and flux control of lipid metabolic pathways. Plant Physiology. 197(2). Article kiae658. https://doi.org/10.1093/plphys/kiae658.
He, Z., Nam, S., Kulkarni, S., Bagheri Kashani, M., Nagarajan, R. 2025. Thermal evaluation of biocomposites made from poly (lactic acid) and cottonseed byproducts. Macromol. 5(2) Article 16. https://doi.org/10.3390/macromol5020016.
He, Z., Nam, S., Tewolde, H., Ford, C.V., Dhandapani, R., Barretto, R., Wang, D. 2025. Morphologic features and thermal characteristics of nine cotton biomass byproducts. Biomass. 5(1). https://doi.org/10.3390/biomass5010012.
Clews, A.T., Whitehead, P.S., Zhang, L., Lu, S., Shockey, J.M., Chapman, K.D., Dyer, J.M., Xu, Y., Mullen, R.T. 2025. Identification and characterization of lipid droplet-associated 2 protein (LDAP) isoforms from tung tree (Vernicia fordii). Plants. 14(5): 814. https://doi.org/10.3390/plants14050814.
Adewumi, O., He, Z., Dhandapani, R., Guo, M. 2025. Lime equivalence values of nineteen biochar products made from defatted cottonseed meal, poultry litter and woody sources for quality assessment. Agricultural & Environmental Letters. 10(1). Article e70017. https://doi.org/10.1002/ael2.70017.
Pundir, A., Thakur, M.S., Prakash, S., Kumari, N., Sharma, N., He, Z., Nam, S., Dhumal, S., Sharma, K., Saxena, S., Kuar, S., Deshmukh, S.V., Kuma, M. 2024. Furfural as a low-volume, high-value asset from agricultural residues: A review on production, agricultural applications and environmental sustainability. Heliyon. 10(15). Article e35077. https://doi.org/10.1016/j.heliyon.2024.e35077.
Zhao, G., Liao, C., Long, H., Cao, H., Zhang, L., 2025. Diacylglycerol Acyltransferase1 Promotes Triacylglycerol Biosynthesis in Oil Tea (Camellia oleifera) Seeds through Transcriptional Activation by WRINKLED1. Journal of Agricultural and Food Chemistry. https://doi.org/10.1021/acs.jafc.5c02120.
Cao, H. 2024. Lipopolysaccharide regulation of antiinflammatory tristetraprolin family and proinflammatory cytokine gene expression in mouse macrophages. BMC Research Notes. 17. Article 82. https://doi.org/10.1186/s13104-024-06743-6.