A Novel Delivery System for the Root Symbiotic Fungus, Sebacina vermifera, and Consequent Biomass Enhancement of Low Lignin COMT Switchgrass Lines
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Sebacina vermifera (MAFF-305830) is a mycorrhizal fungus originally isolated from the roots of orchids that we have previously shown to be tremendously beneficial in enhancing biomass yield and drought tolerance in switchgrass, an important bioenergy crop for cellulosic ethanol production in the United States. Towards this end, we have developed a bentonite clay particle-based delivery system for mass production and dissemination of S. vermifera for large-scale field trials. A greenhouse-based experiment was conducted to evaluate this novel delivery method for biomass enhancement of wild type and transgenic, low lignin (COMT down-regulated) switchgrass lines compared to an efficient in vitro colonization method. S. vermifera colonization enhanced plant biomass regardless of delivery method, although the percentage of fungal biomass in planta increased with the clay-based delivery system. Further, we found that release of some clay minerals in solution was enhanced in the presence of S. vermifera, while others were seemingly reduced. Intriguingly, the presence of S. vermifera has little or no impact on cell wall composition, including lignification. This research is the first report documenting the development of a bentonite clay particle-based delivery system for mass production of any symbiotic microbe and suggests that S. vermifera can be packaged with a mineral composite and effectively delivered to a target host plant.
KeywordsSwitchgrass Mycorrhizae Sebacina COMT
S. vermifera (MAFF-305830) used in this study was obtained from the National Institute of Agro-biological Sciences, Tsukuba, Ibaraki, Japan. The COMT lines used in this study were provided by Chunxiang Fu and Zeng-Yu Wang, Forage Improvement Division, The Samuel Roberts Noble Foundation. We thank Crissa Doeppke, Melissa Glenn, Kimberly Mazza, Logan Schuster, and Kevin Cowley in NREL for their efforts in preparing samples for the HTP recalcitrance pipeline; Erica Gjersing Robert Sykes and Mark Davis in NREL for cell wall composition analysis; David Huhman for ion chromatography; Stacy Allen for qRT-PCR; Jin Nakashima for assistance with SEM and confocal microscopy; Stephen L. Webb for assistance with statistical analysis; and Myoung-Hwan Chi, Blue Stewart, Colleen Elles, and Amanda Hammon for greenhouse assistance. This work was supported by the Bioenergy Science Center, a US Department of Energy Bioenergy Research Center, through the Office of Biological and Environmental Research in the DOE Office of Science.
Conflict of Interest
All the authors in this manuscript declare no conflict of interests inherent to this submission.
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