title: Homologous recombination-mediated gene targeting in the liverwort Marchantia polymorpha L.
abstract: The liverwort Marchantia polymorpha is an emerging model organism on account of its ideal characteristics for molecular genetics in addition to occupying a crucial position in the evolution of land plants. Here we describe a method for gene targeting by applying a positive/negative selection system for reduction of non-homologous random integration to an efficient Agrobacterium-mediated transformation system using M. polymorpha sporelings. The targeting efficiency was evaluated by knocking out the NOP1 gene, which impaired air-chamber formation. Homologous recombination was observed in about 2% of the thalli that passed the positive/negative selection. With the advantage of utilizing the haploid gametophytic generation, this strategy should facilitate further molecular genetic analysis of M. polymorpha, in which many of the mechanisms found in land plants are conserved, yet in a less complex form.
authors.affiliation: Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan.
authors.forename: Kimitsune
authors.initials: K
authors.lastname: Ishizaki
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Johzuka-Hisatomi
authors.forename: Sakiko
authors.initials: S
authors.lastname: Ishida
authors.forename: Shigeru
authors.initials: S
authors.lastname: Iida
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kohchi
mesh_terms: D060054:Agrobacterium / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D005838:Genotype; D059765:Homologous Recombination; D047008:Marchantia / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D018744:DNA, Plant; D008565:Membrane Proteins; D010940:Plant Proteins