Progress 09/01/24 to 08/31/25
Outputs
Target Audience:US undergraduate students with biology and biochemistry majors studying at East Carolina University, HBCUs, and local communities colleges. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The program participants received weekly professional training during the summer research program. Experts from different departments and fields were included to give specific training to these summer research participants, including people from the Office of Environmental Health and Safety, Office of Research Integrity and Compliance, Office of Undergraduate Research, Department of Biology, and the Graduate School. The major professional training included: 1) Lab biosafety; 2) Responsible conduct of research; 3) how to apply for great school; 4) good behavior for undergraduate research. The specific project research training included: 1) plant biotechnology; 2) Agrobacteria-mediated gene transformation; 3) CRISPR/Cas genome editing; and 4) gRNA design and construction. From these trainings, the students not only learned how to do good research and present their discovery but also learn for their future. They also know other research opportunities, such as ECU offering research grants for undergraduate students. These training courses not only trained students for good research, writing papers and doing great presentations but also training them for applying for graduate school and future career. How have the results been disseminated to communities of interest?Presentaions and publications are two major way to diseminate the results from this project. During the second year of this program, the participants made 8 poster presentations during the Summer Undergraduate Research Symposium (July 25, 2025) at East Carolina Campus Piper Hall, Lameesha Calvin, and Ugur Yildiz. Inoculation of Gossypium hirsutum with agrobacteria andpotential to increase parasite resistance Amber Gardner, Stefan Boromisa, Dr. Baohong Zhang. Investigating Cytokinin Morphological Effects on the WUS/CLV3 Feedback Loop in Cotton Shoot Apical Meristems for Enhanced Transformation Parker Ford and Baohong Zhang. Role of MYB Transcription Factor in Fatty Acid Composition of Gossypium hirsutum Mihang Libang ,Nicholas Russell, Isabelle Schenk, Susannah Catherine Webster, Rakesh Kumar PhD, and Baohong Zhang PhD. Knockout of GhPEPC via CRISPR/Cas9 in Gossypium hirsutum Mary Mayone, Dana Kohtz, Dr. Baohong Zhang. Melatonin Priming of Peanut Seeds, Effects on Tolerance to Drought Stress Nicholas Russell, Isabelle Schenk, Susannah Catherine Webster, Mihang Libang, Rakesh Kumar PhD, and Baohong Zhang PhD. Improving Cotton Plant Architecture using CRISPR-Cas9 targeted Editing of Nulliplex branch (GhNB) gene Susannah Catherine Webster, Isabelle Schenk, Mihang Libang, Nicholas Russell, Rakesh Kumar and Baohong Zhang. CRISPR/Cas9-mediated Targeted Editing of GhJAZ Gene for Biotic Stress Tolerance and Fiber Quality Improvement in Cotton Isabelle Schenk, Mihang Libang, Nicholas Russell, Susannah Catherine Webster, Rakesh Kumar, Baohong Zhang. Targeting GhFAD2: Advancing Cotton Transformation via Somatic Embryogenesis An additional poster presentation was accepted for theState ofNorthCarolinaUndergraduate Research & Creativity Symposiumon November 15that Elon University With support from this project, a total of six papers were published in peer-reviewed journals, in which most were with students as co-authors. Zhang Y, Li L, Dai H, Kong X, Rahman M, Zhang B, Zhang Z, Zhou Y, Liu Q: Iron oxide nanoparticles (FeO-NPs) mitigate salt stress in peanut seedlings by enhancing photosynthesis, osmoregulation, and antioxidant activity. Plant Physiology and Biochemistry 2025, 227:110206. Hu N, Tian H, Li Y, Li X, Li D, Li L, Wang S, Zhang Y, Shi X, Huang B, Lu Q, Wang T, Pan X, Tu ., Dai D, Zhang BH, Peng R, Yan F (2025) pHNRhCas9NG, single expression cassette-based dual-component dual-transcription unit CRISPR/Cas9 system for plant genome editing. Trends in Biotechnology, 43(7): 1788-1808. doi: 10.1016/j.tibtech.2025.03.016 Zheng S, Tang Z, Li L, Jia P, Kong X, Xue H, Cheng X, Wang X, Zhang Y, Bai N, Zhang Z, Pan X, Zhang BH (2025) Melatonin reprograms soil microbial community, creates friendly soil environments, and promotes peanut growth. Plant Physiology and Biochemistry 218:109307. doi:https://doi.org/10.1016/j.plaphy.2024.109307 Sun R, Wang Y, Zhu R, Li L, Xi Q, Dai Y, Li J, Cao Y, Guo X, Pan X, Wang Q, Zhang BH (2025) Genome-wide identification of CA genes in cotton and the functional analysis of GhαCA4-D, GhβCA6-D and GhγCA2-D in response to drought and salt stresses. International Journal of Biological Macromolecules 304, 140872. doi: 10.1016/j.ijbiomac.2025.140872 Li L, Cheng X, Zhang Y, Kohtz D, Wang X, Zhang X, Kong X, Xue H, Jia P, Bai N, Li Z, Xiao P, Pan X, Zhang Z, Zhang BH (2024) Exogenous melatonin improves peanut field productivity and quality at reduced nitrogen application. Field Crops Research 319:109650. doi: https://doi.org/10.1016/j.fcr.2024.109650 Li L, Zhang Z, Zhang B (2024) iJAZ: the next breakthrough for engineering pest-resistance in plants? Trends in Plant Science 29 (12):1282-1284. Doi: https://doi.org/10.1016/j.tplants.2024.07.013 What do you plan to do during the next reporting period to accomplish the goals?We already setup website and made an extensive plan for recruiting 10 more students into this program. In 2026 summer, the students will receive great training in in both education and research for developing agricultural workforce for our next generations in the field of agricultural genomics and biotechnology. Hopefully, we will obtain some CRISPR-edited plants for enhancing crop yield, quality and tolerance to various environmental stresses.
Impacts
What was accomplished under these goals?
During the 2nd years, a total of 10 US undergraduate students were recruited into this program and received training in the field of agricultural genomics and biotechnology. During the 10 weeks' research and training, for each week, at least one professional training was given to these students, which included but not limited to 1) Lab biosafety; 2) Responsible conduct of research; 3) how to apply for great school; 4) good behavior for undergraduate research; additionally training also included: 1) plant biotechnology; 2) Agrobacteria-mediated gene transformation; 3) CRISPR/Cas genome editing; and 4) gRNA design and construction. From these trainings, the students not only learned how to do good research and present their discovery but also learn for their future career. The students majorly performed research with 5 different topics :1) cotton genome editing; 2) peanut tissue culture and transgenics; 3) impact of abiotic stresses (drought and salinity) on plant development; 4) impact of nematodes on plant development; and 5) exogenous melatonin impact on peanut growth and development. Based on their results, a total of 8 poster presentations were given by these students. With support from this project, a total of 6 papers were published in peer-reviewed journal, in which most were with students as co-authors.
Publications
- Type:
Other Journal Articles
Status:
Published
Year Published:
2024
Citation:
Li L, Zhang Z, Zhang B (2024) iJAZ: the next breakthrough for engineering pest-resistance in plants? Trends in Plant Science 29 (12):1282-1284. Doi: https://doi.org/10.1016/j.tplants.2024.07.013
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Sun R, Wang Y, Zhu R, Li L, Xi Q, Dai Y, Li J, Cao Y, Guo X, Pan X, Wang Q, Zhang BH (2025) Genome-wide identification of CA genes in cotton and the functional analysis of Gh?CA4-D, Gh?CA6-D and Gh?CA2-D in response to drought and salt stresses. International Journal of Biological Macromolecules 304, 140872. doi: 10.1016/j.ijbiomac.2025.140872
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Zheng S, Tang Z, Li L, Jia P, Kong X, Xue H, Cheng X, Wang X, Zhang Y, Bai N, Zhang Z, Pan X, Zhang BH (2025) Melatonin reprograms soil microbial community, creates friendly soil environments, and promotes peanut growth. Plant Physiology and Biochemistry 218:109307. doi:https://doi.org/10.1016/j.plaphy.2024.109307
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Hu N, Tian H, Li Y, Li X, Li D, Li L, Wang S, Zhang Y, Shi X, Huang B, Lu Q, Wang T, Pan X, Tu ., Dai D, Zhang BH, Peng R, Yan F (2025) pHNRhCas9NG, single expression cassette-based dual-component dual-transcription unit CRISPR/Cas9 system for plant genome editing. Trends in Biotechnology, 43(7): 1788-1808. doi: 10.1016/j.tibtech.2025.03.016
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Zhang Y, Li L, Dai H, Kong X, Rahman M, Zhang B, Zhang Z, Zhou Y, Liu Q: Iron oxide nanoparticles (FeO-NPs) mitigate salt stress in peanut seedlings by enhancing photosynthesis, osmoregulation, and antioxidant activity. Plant Physiology and Biochemistry 2025, 227:110206.
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Progress 09/01/23 to 08/31/24
Outputs
Target Audience:
Nothing Reported
Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?1. Student Training The program participants received weekly professional training during the summer research program. Experts from different departments and fields were included to give specific training to these summer research participants, including people from the Office of Environmental Health and Safety, Office of Research Integrity and Compliance, Office of Undergraduate Research, Department of Biology, and the Graduate School. The major professional training included: 1) Lab biosafety; 2) Responsible conduct of research; 3) how to apply for great school; 4) good behavior for undergraduate research. The specific project research training included: 1) plant stress physiology; 2) CRISPR/Cas genome editing; 3) gRNA design and construction; and 4) plant transformation and transgenics. From this training, the students not only learned how to do good research and present their discovery but also learn for their future. They also know other research opportunities, such as ECU offering research grants for undergraduate students. These training courses not only trained students for good research, writing papers and doing great presentations but also training them for applying for graduate school and future career. 2. Student Outreach To enhance the students' social activities and research interest, the students have performed the following social and professional activities: Attending the Cotton Breeders Meeting at Raleigh, North Carolina During the 2024 summer research program, the student participants got a chance to attend the Cotton Breeders meeting organized by the Cotton Incorporated in Raleigh, North Carolina. During this meeting, the participants attended seminars and talked with scientists in the field of cotton genomics and biotechnology. Attending this meeting inspired all the participants to do great research in the field of plant genomics and biotechnology. Visiting Dule University and Wright Brothers National Memorial During this program, we used two Saturdays to visit Duke University and Wright Brothers National Memorial. During these two visits, the participants made great interaction with other students and staff and learned something outside of this program, which may inspire them to better science and attend a higher level of educations, such as attending graduate school. 3. Student Research To train undergraduate student research for their career and our next generation workforce, by learning first-hand research, is one of the major goals for this summer program. To achieve this goal, the 10 students were divided into 5 groups to perform the following 5 different projects: 1) cotton genome editing; 2) peanut tissue culture; 3) impact of nanoparticles on plant development; 4) exogenous melatonin impact on peanut growth and development. Cotton genome editing and biotechnology CRISPR/Cas9 genome editing is a reliable and powerful biotechnical tool for precisely editing a specific gene at a specific location. Since it was inverted in 2012, CRISPR/Cas9 genome editing has been widely used in gene functional study and crop improvement for enhancing crop yield, quality and tolerance to various environmental stresses. In this 10-weeks summer program, all 10 participants (5 major and 5 minor) have performed research in this field, in which they employed CRISPR/Cas9 to target genes associated with oil biosynthesis. The participants designed two gRNAs for each of 6 selected targeted genes and cloned the gRNAs into vectors and transferred into E. coli and Agrobacteria. Finally, they employed Agrobacteria-mediated transformation delivered the gRNAs and Cas9 protein gene into cotton cells. 2) Peanut tissue culture Peanuts are a vital crop for their edible seeds and oil production. Creating an efficient peanut regeneration system is important for the regeneration of transgenic peanut plants. This study's objective was to find effective methods for regenerating Arachis hypogaea L. (peanut) plants through different concentrations and mixtures of plant growth hormones. The peanut cotyledons were used as the explants and were cultured on four different hormone mixtures for shoot regeneration media. Following six weeks, the shoot regeneration was recorded. After the shoots were regenerated, they were cultured on four different hormone concentrations and mixtures for root regeneration media and four weeks later the root number, root length, shoot length, and leaf number were recorded. On the best medium, each explants generated average of 9.1 and 7.7 shoots respectively with root length of 4.2 inches and shoot length of 3.5 inches. After testing different factors, including explants, genotypes and different plant growth regulators, high efficient plant regeneration was obtained from peanut explants; the plant regeneration ratios reached 90%. This provides foundation for obtaining transgenic peanuts and genome editing for crop improvement. 3) Impact of nanoparticles on plant development Cotton (Gossypium hirsutum L.) plays an important role in many different aspects, such as the textile industry, industrial products, agriculture, and science. Finding effective growth techniques for cotton is crucial to maintaining high-quality production, providing a steady supply, and creating different resilient strains that can withstand disease, unfavorable climate conditions, and pests. This study observes the effects of different concentrations of nanoparticles on the roots and leaves of cotton plants. Our hypothesis is that lower concentrations of nanoparticles will promote plant growth while higher amounts of nanoparticles will inhibit growth. To test this hypothesis, cotton seedlings cultured in hydroponic boxes were treated with different amounts of nano-aluminum oxide (0, 50, 100 and 200 mg/L), and the biomass, malonaldehyde (MDA) content, soluble protein (SP), activity of superoxide dismutase (SOD), guaiacol-peroxidase (G-POD), and catalase (CAT) were determined. Results varied for each of the tests due to the different concentrations of nanoparticles. As the concentration of aluminum oxide increased, the dry weight of the shoot and root increased first and then decreased, and the highest values were obtained in plants treated with 100 mg/L nano-aluminum oxide. With the increase in the concentration of aluminum oxide, MDA content showed a decreased and then an increased trend both in leaves and roots. The SP content had an increased trend in the leaves and a decreased trend in roots. The activity of SOD and POD had an increased and decreased trend in the leaves and an increased trend in the roots; the CAT activity had an increased trend in the leaves and an increased and decreased in roots. In conclusion, suitable concentration of nano-aluminum oxide could promote the growth of cotton seedlings and enhance the activity of the antioxidant enzyme such as SOD, G-POD and CAT, and decrease the degree of membrane peroxidation 4) Exogenous melatonin impact on peanut growth and development In this study, we studied the impact of melatonin application on peanut growth and development at the reduced application of nitrogen (N). Melatonin-treated plants had more stable carbon and N metabolism than the untreated ones. This stability was linked to improved photosynthesis, sucrose production, and N assimilation, especially at the reduced N levels (90 and 135?kg/ha). Across three N levels and two years of field tests, MT increased peanut dry matter by 23.49?% from 455.63?g/m2to 562.66?g/m2, enhanced the accumulation and mobilization of dry matter and N to grains by increasing peanut grain mass by 22.41-29.07?% at different N levels. This process appears to subsequently elevate the effective pod rate, leading to an average increase in pod yield, fat and protein content by 12.63?%, 7.95?%, and 10.33?%, respectively, over a two-year period and across three N application levels. How have the results been disseminated to communities of interest?During the 1st year of this program, the participants made seven poster presentations, which included six during the Summer Undergraduate Research Symposium (July 26, 2024 ) at East Carolina Campus Leslie Mendoza, Dana Kohtz, Dr. Lijie Li, Dr. Baohong Zhang. Effects of Different Concentrations of Nano-Aluminum on Antioxidant Enzyme Activity of Cotton Seedlings Dana Kohtz, Leslie Mendoza2, Dr. Lijie Li, Dr. Vijay Sheri, Dr. Baohong Zhang. Establishment of an Efficient Peanut Regeneration System Stefan Boromisa, Zachary Myers, Parker Ford. Optimization of Shoot Multiplication Media in Elite Cotton Cultivars Ruben Garcia, Vijay Sheri, and Baohong Zhang. Modification TCP4 Transcription Factor Role in Fatty Acid Composition of Gossypium hirsutum Holmes Luke Anthony,Vijay Sheri, and Baohong Zhang. Gene Editing of PEPC2 Gene in Gossypium hirsutum using an Agrobacterium Transformation Method Linda Xue, Vijay Sheri, Baohong Zhang. MYB-89 TF for Increased FA Production and Oil Yield in Gossypium hirsutum Yousif Sinan Sousan, Vijay Sheri, and Baohong Zhang. Gene Editing ofGossypium hirsutum UsingAgrobacteriumT.MediatedTransformation An additional poster presentation was accepted for theAnnual Biomedical Research Conference for Minoritized Scientists (ABRCMS) 2024 at Pittsburgh, PA on November 13-16. Leslie Mendoza, Dana Kohtz, Dr. Lijie Li, Dr. Baohong Zhang. Effects of Different Concentrations of Nano-Aluminum on Antioxidant Enzyme Activity of Cotton Seedlings What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, 10 undergraduate students will be recruited and selected into this program to perform and learn the first hand research in the field of plant genome and agricultural biotechnology. Based on the students' research, at least two publications and 5 poster presentation will be generated in the 2nd year of this project.
Impacts
What was accomplished under these goals?
During the first year of this program, 10 undergraduate students were selected and participated in this program from a total of 33 completed applications. These participants came from East Caolina University, HBCU Fayetteville State University and local community colleges (Pitt Community College). The selected students performed 10 weeks' research and training in the field of plant genomics and biotechnology at East Carolina Campus. For each week, at least one professional training was given to these students, which included but not limited to 1) Lab biosafety; 2) Responsible conduct of research; 3) how to apply for great school; 4) good behavior for undergraduate research; additionally training also included: 1) plant stress physiology; 2) CRISPR/Cas genome editing; 3) gRNA design and construction; and 4) plant transformation and transgenics. From this training, the students not only learned how to do good research and present their discovery but also learn for their future. The students also get a chance to attend one international conference on Cotton Genome and Breeding Tours in Raleigh, NC organized by the Cotton Incorporated. The students majorly performed research with 5 different topics :1) cotton genome editing; 2) peanut tissue culture; 3) impact of nanoparticles on plant development; 4) impact of nematodes on plant development; and 5) exogenous melatonin impact on peanut growth and development. Based on their results, a total of 7 poster presentations were given by these students.
Publications
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