Improving Bioenergy Lignocellulosic Feedstock Through CRISPR-Cas9 Technology in Switchgrass (Panicum Virgatum L.)
Increasing greenhouse gas emissions (GHG) due to the use of fossil fuels has a negative impact on the climate, with an estimated increase in global temperature of 6°C if no action is taken. To combat this increasing GHG, particularly Carbon dioxide, the use of biofuels as a replacement for petroleum-based fuels in transportation has been proposed. Biofuels, energy sources generated from biomass have emerged as a potential route to meet energy demand and avoid political instability and environmental issues worldwide. Switchgrass (Panicum virgatum L.), a native North America grassland species has been considered as an excellent bioenergy feedstock due to its high biomass yield, wide adaptation, as well as its lower input energy for production. The major limitation of using switchgrass for biofuels is (i) the difficulty to degrade the main constituents of cell wall biomass, which in some instances is due to its recalcitrance behavior, (ii) to practice convectional breeding as the grass is self-incompatible and has varied ploidy levels. This fully obstacles the exploitation of this crop as a lignocellulosic biofuel. This creates needs for the development of genetic manipulation strategies to produce improved biomass. To achieve this, high-throughput efficient in-vitro cell culture systems capable of genetic transformation and regeneration are necessary for studying gene functions and germplasm improvement. In this study we developed reliable transformation and regeneration methods for cloning biosynthetic lignin cell wall genes with CRISPR-Cas9 technology, which provides an avenue for gene editing and crop improvement, thus leading to a better understanding of lignin functions in switchgrass biomass recalcitrance and at the same time provide fast and reliable methods to alter cell wall biosynthesis and improve biomass digestibility/conversion of this crop. Currently, we established embryogenic cell cultures from friable callus-derived seeds or inflorescences of Alamo and P32. Established cell lines are being used for transformation with subsequent plant regeneration for cell wall genes editing
Plant sciences|Agricultural chemistry|Alternative Energy
Wilson Kihugu Ouma,
"Improving Bioenergy Lignocellulosic Feedstock Through CRISPR-Cas9 Technology in Switchgrass (Panicum Virgatum L.)"
ETD Collection for Tennessee State University.