Discipline: Biological Sciences
Subcategory: Plant Research
Room: Exhibit Hall
Dakota Walker - Fort Valley State University
Co-Author(s): Heather Gladfelter- University of Georgia, Athens, Ga; Dr. Dayton Wilde- University of Georgia, Athens, Ga
Bioengineering of Ornamental Plants Through Organogenesis and Somatic EmbryogenesisHighly prized ornamental species, Cornus florida, Gerbera jamesonii, and Rosa hybrida face challenges from fungal and viral pathogens. The focus of this study was to develop genetic engineering protocols to confer resistance to their respective ornamental pest. For Cornus florida, three somatic embryogenic lines were transformed using Agrobacterium strains/binary plamid: AGL1+GUSi and LBA4404+GFP. Cornus flordia is placed on a liquid selection of Geneticin 4 mg/L+ WPM media + 0.1 IBA+ 300 mg/L timentin for 4 weeks on rotator shaker at 100 rmp in the dark. After 5 weeks, the cells are placed onto nyloned semi-solid selection media. Cornus flordia demonstrated a successful transformation via GFP and GUSi expression in putative transgenic SE cells. For Gerbera jamesonii, leaf explants were Agro-infected with binary plasmid containing C58+GFP. After initial infection, the explants are placed on a semi-solid selection of Hygromycin 0,10,15, and 20 mg/L+ MS media +0.1mg/L IBA+2mg/L BAP+ 400 mg/L timentin in the light. Via fluorescent microscopy, the explants are studied for GFP expression after a 14-day subculture rotation to fresh selection media. The result of which demonstrate a successful transformation via GFP in putative transgenic leaves. Lastly, for Rosa hybrida, different media compositions were tested for in vitro regeneration method using young leaf explants. Young leaves below the apex of the in vitro shoot clump are placed on callus induction MS media+2.5 mg/L 2,4-D with an induction period for 30 days in the dark at room temperature (23+/- ◦C). The results of which allows for callus formation on leaf petioles and mid-vein of leaf. In future research, after callus has accumulated, the callus is placed on SE media, ½ MS+ 1 mg/L GA3+0.5 mg/L TDZ in the light for 60 days at room temperature. The long-term goal is to use CRISPR technology to confer resistance to powdery mildew, a devastating fungal pathogen to Cornus floridaand RNA interference (RNAi) technology to confer resistance to rose rosette virus to Rosa hybrida. References: Korban, S.S. (2005). Somatic embryogenesis in Rose: Gene expression and genetic transformation. Plant Cell Monogr (2). In: Somatic Embryogenesis (Ed. Mujib, A and Samaj, J.), Springer-Verlag Berlin, Heidelberg: 247-257.Lloyd, G. and McCown, B. (1980). Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Proc. Intern. Plant Prop. Soc., 30: 421-437.Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15: 473-497.
Funder Acknowledgement(s): Funder Acknowledgement(s): This study was supported, in part, by a grant from UGAFVSU The Rising Scholars Internship program. The author would like to thank for financial support from NSF HBCU-UP (HRD-2011903), S-STEM (DUE-1834046), and the Department of Education MSEIP (P120A2000016) programs at Fort Valley State University.
Faculty Advisor: Dr. Sarwan Dhir, email@example.com
Role: I prepared the media, explants, agrobacteria, subcultures, and collected the data. After conducting the experiments, I did the analyzation, presentation preparation and interpretation of results. The process was supervised by Dr. Heather Gladfelter and Dr. Dayton Wilde.