MISeqv2 of Seed fungal endophyte: Switchgrass
Seeds were Cave-in-Rock switchgrass variety from 2007 lot SFD-07-F11 (USDA Elsberry Plant Materials Center, Elsberry, MO). Seeds were surface sterilized and stratified at 4C in petri dishes with autoclave filter paper soaked with nanopure water for ~2 months. On July 10, 2018, a random subset of five groups of 3-5 seeds were frozen at -80C for sequencing to characterize the putative maternal community. plant samples were surface sterilized by placing material (either seed or leaf) in a tissue cassette and submerging the sample in a series of 1min in nanopure water, 1 minute in 70% ethanol, 5 min in 10% bleach, and 1 min in 90% ethanol [40]. Plant material was extracted using Plant DNeasy kit with standard manufacture protocol and an extended lyses step. We amplified the ITS2 region using the fungal specific primers ITS9 (GAACGCAGCRAAIIGYGA; [41]) and ITS4 (TCCTCCGCTTATTGATATGC [42]).