Progress 04/15/24 to 04/14/25
Outputs
Target Audience:Undergraduate and graduate students, plant biology researchers in academia, and scientists in biotechnology industries focus on oilseed improvement, photosynthesis, and renewable energy. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided a training opportunity for undergraduate and graduate students during the reporting period. How have the results been disseminated to communities of interest?Publications Deo, B., Muthan, B., Cruise, T.L.A., Mukherjee, T., Allen, D.K., & Sanjaya, S.A. (2025). Flue Gas Desulfurized Gypsum as a Sustainable Amendment for Coal Mine Soil Reclamation and Bioenergy Crop Production. Science of the Total Environment (In Review). Sanjaya, S. (2025). How biotechnology can revolutionize agriculture: feeding the world and powering the future in the face of climate change. In proceedings of an International Conference on Future perspectives in plant science (ICFPPS-2025). PSG College of Arts and Science, Coimbatore, India. pp-24. OSBN 978-81-980915-0-5. Poster Presentations: Muthan, B., Khatiwada, S., and Sanjaya, S. (2024). Application of Gene Technology for Increasing Oil in Bioenergy Crops. Midwest ASPB meeting, Purdue University, March 2024. Muthan, B., Khatiwada, S., Jayaraman, K and Sanju Sanjaya (2024). Genetic engineering of switchgrass biomass for bioenergy production. ASPB meeting, Honolulu, Hawaii, June 2024. Bagyalakshmi Muthan, Jie Wang, Ruth Welti, Dylan K. Kosma, Linhui Yu, Bikash Deo, Subhiksha Khatiwada, Vijaya K.R. Vulavala, Kevin L. Childs, Changcheng Xu, Timothy P. Durrett, Sanju A. Sanjaya (2024). How Aquatic Plants Develop Tolerance to Heavy Metal Toxicity. 26th International Symposium On Plant Lipids, UNL, July 2024. Oral Presentations: Sanjaya, S. (2024). Understanding How Plants Tolerate Heavy Metal Stress: The molecular Mechanisms. East Tennessee University, TN, USA. Sanjaya, S. (2025). Lipid metabolism in duckweed under heavy metal stress conditions. Gordon Research Conference on Plant Lipids: Structure, metabolism and functions, Pomona, CA, USA. Sanjaya, S. (2025). Molecular mechanisms of heavy metal stress tolerance in Spirodela polyrhiza. Plant and Animal Genome Conference 32 (PAG32), San Diego, CA, USA. Sanjaya, S. (2025). Molecular Mechanisms of Plant Tolerance to Heavy Metal Stress. North Carolina A & T State University, Greensboro, NC, USA. Sanjaya, S. (2025). Genetic Transformation of Recalcitrant Upland Energy Crop Switchgrass for Enhanced Oil Yield. Plant and Animal Genome Conference, Asia, New Delhi, India. Sanjaya, S. (2025). How biotechnology can revolutionize agriculture: feeding the world and powering the future in the face of climate change. PSG College of Arts and Science, Coimbatore, India. Sanjaya, S. (2025). Potentials of Biotechnology to Produce Renewable Hydrocarbons in Plants for Dealing with the Effect of Climate Change. University of Mysore, India. Sanjaya, S. (2025). Metabolic engineering of hydrocarbons in plants for bioenergy production. PES University, Bangalore, India Sanjaya, S. (2025). Can the application of biotechnology techniques help to feed and fuel the world? M.S. Ramaiah College of Arts, Science & Commerce, Bangalore, India. What do you plan to do during the next reporting period to accomplish the goals?We will continue comprehensive analyses of key primary and storage metabolites across all mutant and transgenic lines of Arabidopsis thaliana and Camelina sativa. Parallel efforts are underway for seed bulking in preparation for controlled-environment and field performance trials. Additional work will include genotyping of transgenic events and detailed assessment of growth and yield parameters, including photosynthetic performance and seed metabolite profiles. Furthermore, we will advance our genetic interaction studies through transgenic approaches to elucidate the functional interplay between WRI1 and associated metabolic pathways involved in carbon allocation and storage.
Impacts
What was accomplished under these goals?
The temporal expression analysis of triose transporters and WRI1 transcription factor in Arabidopsis thaliana siliques revealed a dynamic regulatory relationship during seed development. WRI1 transcript levels increased markedly from 5 days after pollination (DAP) and remained consistently elevated through 9 DAP. This upregulation corresponded with a significant induction of its downstream targets, indicating transcriptional activation mediated by WRI1 in reproductive tissues independent of tissue specificity. Concurrently, a substantial and continuous decline in free glucose content was observed from 3 to 15 DAP. This inverse relationship suggests active transport of substrates into plastids, where they are likely utilized in the biosynthesis of storage compounds such as starch and triacylglycerols (TAGs). To further investigate this regulatory mechanism and its impact on carbon assimilation, we have generated a series of transgenic Arabidopsis lines overexpressing trisphosphate transporters individually or in combination with Arabidopsis WRI transcription factor, driven by constitutive and seed-specific promoters and compared with wild-type and mutant lines. Arabidopsis transgenic lines exhibited enhanced net CO? assimilation rates under increasing light intensity compared to wild types. Notably, trisphosphate transporter lines, particularly those in combination with WRI1, showed improved assimilation under high light. In contrast, all mutant lines displayed significantly reduced photosynthetic performance. The double mutant showed early light saturation and poor assimilation under low light, highlighting a synergistic role by these genes in photosynthetic acclimation. CO? response analysis revealed reduced net assimilation in the double mutant across optimal and suboptimal internal CO? concentrations. This suggests a critical regulatory role of these genes in sustaining CO? assimilation capacity, possibly through its influence on key metabolic and transport pathways. Building upon these promising results, we have advanced to translational studies in Camelina sativa, an oilseed crop with bioenergy potential. T3 generation transgenic camelina lines overexpressing triose transporters, either independently or in combination with WRI1 transcription factor under the control of constitutive and seed specific promoters, respectively. Ectopic expression of these transporter genes significantly enhances photosynthetic efficiency of transgenic plants during the vegetative growth phase. The transgenic plants exhibited higher CO? assimilation under both increasing light intensities and ambient CO? concentrations. These improvements were associated with enhanced biochemical factors such as Vcmax, Jmax, and TPU, which contribute to better photosynthetic performance. Furthermore, our studies revealed the increased CO? assimilation in siliques and improved seed yield and seed morphology in transgenic plants, demonstrating the potential for enhanced carbon partitioning towards seeds, thus increasing seed yield. Experiments on the quantification of seed storage metabolites in T3 generation seeds and the bulking of seeds for field evaluation are underway.
Publications
- Type:
Other Journal Articles
Status:
Under Review
Year Published:
2025
Citation:
" Deo, B., Muthan, B., Cruise, T.L.A., Mukherjee, T., Allen, D.K., & Sanjaya, S.A. (2025). Flue Gas Desulfurized Gypsum as a Sustainable Amendment for Coal Mine Soil Reclamation and Bioenergy Crop Production. Science of the Total Environment (In Review).
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
" Sanjaya, S. (2025). How biotechnology can revolutionize agriculture: feeding the world and powering the future in the face of climate change. In proceedings of an International Conference on Future perspectives in plant science (ICFPPS-2025). PSG College of Arts and Science, Coimbatore, India. pp-24. OSBN 978-81-980915-0-5.
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Progress 04/15/23 to 04/14/24
Outputs
Target Audience:The research participants comprised undergraduate and graduate students, visiting scholars, academic plant biology researchers, scientists from biotechnology industries focused on food and bioenergy, local non-governmental organizations, and farmers. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided training opportunities for undergraduatestudents during the reporting period. Project outcomes were also formally and informally shared with scientists and farmers focused on agriculture, plant biology, food and renewable energy. How have the results been disseminated to communities of interest?Poster presentations: Muthan, B., Khatiwada, S., and Sanjaya, S. (2024). Application of Gene Technology for Increasing Oil in Bioenergy Crops. Midwest ASPB meeting, Purdue University, 2024. Muthan, B., Khatiwada, S., Jayaraman, K and Sanju Sanjaya (2024). Increasing energy density of switchgrass biomass for bioenergy production. In the 21st Research Symposium of the Association of 1890 Research Directors. Nashville, TN, April 2024. Cole, G., Patil, S., Muthan, B., and Sanjaya, S. (2024). Understanding the Effect of Carbohydrates on Seed Germination and Establishment in Arabidopsis. In the 21st Research Symposium of the Association of 1890 Research Directors. Nashville, TN, April 2024. Oral presentations: Sanjaya, S. (2024). Climate Change and Biotechnology for Improving Production of Food and Fuels. In the 21st Research Symposium of the Association of 1890 Research Directors. Nashville, TN, April 2024. Sanjaya. Molecular mechanisms of heavy metal stress tolerance in plants. Kansas State University, February 2024. Sanjaya. Can we meet the world's growing demand for food and energy using biotechnology, University of Tennessee, April 2023. Sanjaya. Potentials of Biotechnology to Produce Renewable Hydrocarbons in Plants for Dealing with the Effect of Climate Change. New frontiers of natural sciences: 2nd international selçuk meeting October 02-04, 2023 Konya-Türkiye Sanjay. Energy and environmental science research and educational opportunities and challenges in West Virginia, University of Tennessee, October 2023. What do you plan to do during the next reporting period to accomplish the goals?We will continue analyzing candidate genes identified by our RNA-Seq analysis using forward and reverse genetic approaches. Additionally, we will establish the molecular and biochemical mechanisms regulating carbon partitioning into oils using mutant and transgenic lines. Bulk seed genotyping and growth yield measurements, including photosynthesis rates and seed metabolites, will be performed. Finally, we will continue genetic interaction studies using both in vivo and in vitro methods.
Impacts
What was accomplished under these goals?
We utilized bioinformatics tools to analyze RNA-Seq data. Low-quality, short, prematurely terminated, and ambiguous base reads were removed from the raw sequences. The remaining clean reads were aligned to the Arabidopsis genome assembly TAIR10 using TopHat and BWA software. Comparative transcriptomics of sucrose-treated Arabidopsis mutants, transgenics, and wild-type Col-2 plants revealed altered expression of over 400 genes involved in central carbon metabolism, including photosynthesis, glycolysis, fatty acid metabolism, triacylglycerol synthesis, starch assembly, and transport. Sucrose also downregulated multiple lipase genes in mutants and transgenics, potentially impacting energy production and nutrient acquisition. Further bioinformatics experiments are underway to elucidate the functions of unknown genes. We have obtained a purified antibody against the target transcription factor and confirmed its efficacy via western blot. Pull-down assays are in progress to identify promoter binding sites that regulate oil biosynthesis. Using a LI-6800 plant chamber, we measured CO2 response curves in 15-day-old Arabidopsis seedlings within 5 hours of the light cycle onset at 800 μmol constant airflow. Preliminary data showed higher CO2 assimilation in transgenic versus mutant and control plants. Light response curves using a LiCOR 6800 with leaf cuvettes and a fluorometer revealed increased photosynthetic capacity in transgenics compared to mutants. Measurements were taken at 1500 μmol photon m-2 s-1 irradiance, 400 μmol CO2 mol-1, and 23°C leaf temperature. We have initiated greenhouse studies on transgenic camelina lines to evaluate agronomic performance and seed yield and composition.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Muthan, B., Wang, J., Welti, R., Kosma, D. K., Yu, L., Deo, B., Khatiwada, S., Vulavala, K.R. V., Childs, L.K., Xu, C., Durrett, P.D., & Sanjaya, S. A. (2024). Mechanisms of Spirodela polyrhiza Tolerance to FGD Wastewater-Induced Heavy-Metal Stress: Lipidomics, Transcriptomics, and Functional Validation. Journal of Hazardous Materials, 133951.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Sanjaya, S. (2024). Climate Change and Biotechnology for Improving the Production of Food and Fuels. Pp 265. In the 21st Research Symposium of the Association of 1890 Research Directors. Nashville, TN, April 2024.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Cole, G., Patil, S., Muthan, B., and Sanjaya, S. (2024). Understanding the Effect of Carbohydrates on Seed Germination and Establishment in Arabidopsis. Pp 429-430. In the 21st Research Symposium of the Association of 1890 Research Directors. Nashville, TN, April 2024.
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