Hornwort (Anthoceros agrestis) biolistics-mediated transformation protocol v1

Here we report an efficient biolistics method for generating transient-expression and stable transgenic lines in the model hornwort,Anthoceros agrestis. An average of 569 (± 268) cells showed transient expression per bombardment, with green fluorescent protein expression observed within 48 hours. A total of 81 stably transformed lines were recovered across three separate experiments, averaging six lines per bombardment. We followed the same method to transiently transform nine additional hornwort species, and obtained stable transformants from A. fusiformis.