Student/presenter: Chidinma Lois Nwoko, MS student, Plant and Soil Science
Format: Poster presentation
Title: Development of an efficient method for protoplast isolation, transfection, and gene editing form soybean roots
Chidinma Lois Nwoko, Arjun Ojha, Vikas Devkar, Gunvant B. Patil
Institute of Genomics for Crop Abiotic Stress Tolerance (IGCAST), Department of Plant & Soil Science, Texas Tech University, Lubbock, TX
Protoplasts are plant cells with degraded cell wall that behave like animal cells in vitro. Protoplast is a versatile system in modern plant biology that provides a platform for rapid analysis of diverse signaling pathways, studying functions of cellular machineries and functional genomics screening. Protoplast allows the direct delivery of DNA, RNA or protein into the plant cell and provides a high-throughput system to validate gene-editing reagents. However, this system is less exploited in several legumes crops (including soybean), and it is because of lower protoplast yields, transfection efficiencies and lack of working protocol for plant regeneration from protoplast Moreover, protoplast isolation in several plant is mainly focused on leaf mesophyll tissues. Although, root tissues provide several advantages, root protoplast isolation, transfection and geneediting have not been established in soybean. To overcome these bottlenecks, we are developing a new robust method for high quality protoplast isolation and transfection from soybean roots (including transgenic hairy roots). With our newly developed the highest yield, 1.3 x 106 and 7.3 x 105) of protoplasts were obtained from soybean roots and hairy roots respectively. More importantly, we also describe a method for gene-editing in soybean protoplasts isolated from root tissues.