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An Agrobacterium ‐mediated stable transformation technique for the hornwort model Anthoceros agrestis
Author(s) -
Frangedakis Eftychios,
Waller Manuel,
Nishiyama Tomoaki,
Tsukaya Hirokazu,
Xu Xia,
Yue Yuling,
Tjahjadi Michelle,
Gunadi Andika,
Van Eck Joyce,
Li FayWei,
Szövényi Péter,
Sakakibara Keiko
Publication year - 2021
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/nph.17524
Subject(s) - biology , botany , agrobacterium , transformation (genetics) , genetics , gene
Summary Despite their key phylogenetic position and their unique biology, hornworts have been widely overlooked. Until recently there was no hornwort model species amenable to systematic experimental investigation. Anthoceros agrestis has been proposed as the model species to study hornwort biology. We have developed an Agrobacterium ‐mediated method for the stable transformation of A. agrestis , a hornwort model species for which a genetic manipulation technique was not yet available. High transformation efficiency was achieved by using thallus tissue grown under low light conditions. We generated a total of 274 transgenic A. agrestis lines expressing the β‐glucuronidase (GUS), cyan, green, and yellow fluorescent proteins under control of the CaMV 35S promoter and several endogenous promoters. Nuclear and plasma membrane localization with multiple color fluorescent proteins was also confirmed. The transformation technique described here should pave the way for detailed molecular and genetic studies of hornwort biology, providing much needed insight into the molecular mechanisms underlying symbiosis, carbon‐concentrating mechanism, RNA editing and land plant evolution in general.