Utilizing Beneficial Bacterial Endophytes to Promote Switchgrass Growth in Low- input Agricultural Production Systems

The US Department of Energy has focused research efforts on developing switchgrass into a bioenergy feedstock, helping to offset the use of non-renewable fossil fuels and make the US more energy independent. Bacterial endophytes, which reside inside plant tissues, are proven to increase yield and stress resistance in a number of plants. The primary objective of this dissertation was to explore the use of endophytes to improve biomass yields of switchgrass on lands not suitable for food crops and better understand the underlying mechanisms of the plant-endophyte interaction. Integration of this research into K-12 STEM education to increase interest in plant sciences and create the next generation of scientists with the motivation to help solve the challenges facing society in the twenty first century was the objective of the outreach component of this project. Chapter one demonstrates the ability of Burkholderia phytofirmans strain PsJN to colonize switchgrass and promote plant growth under in vitro (approximately 50% higher), and growth chamber and greenhouse (48.6% higher biomass yields) conditions. The objectives of Chapter two were to determine stand establishment in the field with different nutrient levels. PsJN bacterization positively benefited growth and development of switchgrass seedlings in the field with both low and high nutrient content. Highly significant (p<0.001) stimulation of root and shoot growth, lateral root formation and number of tillers was recorded on soil with low fertility. PsJN bacterization also enhanced biomass accumulation during the two seasons of growth on both poor (p<0.001) and rich (p<0.05) soil, indicating the potential for the use of PsJN in a low-input switchgrass feedstock production system. Chapter three outlines differences in gene expression patterns upon bacterization, between the responsive cv. Alamo, and a non-responsive cv. Cave-in-Rock. Using EST microarrays and quantitative PCR up- and down-regulated genes were identified in both cultivars. One of the key genes identified was a member of the tau class, glutathione S-transferase (GST). GST enzymes are known to be involved in plants responses to stress. Using overexpression and knockout/knockdown techniques we demonstrated that GST is likely involved in the bacterization induced early plant growth promotion in switchgrass. Chapter four describes the potential for the utilization of beneficial bacterial endophytes capable of fixing atmospheric nitrogen in a free-living state in the development of low-input switchgrass feedstock production systems. Sphingomonas sp. strain NSL isolated from switchgrass tissue was able to grow on nitrogen free medium and stimulated growth of switchgrass cv. Alamo under nitrogen deficient conditions. The ability to fix atmospheric nitrogen was also moved to Burkholderia phytofirmans strain PsJN via horizontal gene transfer from the legume nodulating Burkholderia phymatum. The transformed PsJN was able to fix nitrogen and promote plant growth under nitrogen limited conditions. At every step of the research described in this dissertation efforts were made to include its elements into K-12 education. Chapter five describes four case studies aiming at the enhancement of youth interest in plant sciences in the socieoeconomically depressed areas of Southside Virginia.