Student/presenter: Nwoko Chidinma Lois, Msc. Student, Crop Molecular Improvement Research
Format: Oral presentation
Title: An improved method for protoplast isolation and gene-editing from soybean root, callus and transgenic hairy-roots
Chidinma Lois Nwoko, Arjun Ojha Kshetry, Vikas Devkar and Gunvant B. Patil
Institute of Genomics for Crop Abiotic Stress Tolerance, Department of Plant and Soil Sciences, Texas Tech University, Lubbock, TX, 79409, USA
Protoplasts are a useful system in plant biology because they provide a platform for the rapid evaluation of a variety of signaling pathways, the investigation of gene function, as well as a high-throughput instrument for the study of functional genomics. This makes protoplasts an important component of the study of plant biology. This technique is less explored in various crops, the most notable of which is soybean, because it is difficult to isolate high-quality protoplast from a varied variety of plant tissues. While isolating protoplasts from leaf mesophyll is the most common method, isolating them from other tissues can provide researchers with other alternatives and provide a wider range of cells for tissue-specific experiments like single-cell transcriptomics. In addition to this, it helps to ensure that the cells that have been gathered are a representation of the plant as a whole and not just of a certain tissue. In conclusion, the isolation of protoplast from transgenic tissues, such as hairy roots and transgenic callus, is a quick method that may be used to assess gene activity. To facilitate future regeneration, we created a streamlined method of isolating, transfecting, and editing genes in both transgenic (hairy-roots and callus) and non-transgenic (roots) protoplasts.