title: An efficient reverse genetics platform in the model legume Medicago truncatula.
abstract: Medicago truncatula is one of the model species for legume studies. In an effort to develop legume genetics resources, > 21 700 Tnt1 retrotransposon insertion lines have been generated. To facilitate fast-growing needs in functional genomics, two reverse genetics approaches have been established: web-based database searching and PCR-based reverse screening. More than 840 genes have been reverse screened using the PCR-based approach over the past 6 yr to identify mutants in these genes. Overall, c. 84% (705 genes) success rate was achieved in identifying mutants with at least one Tnt1 insertion, of which c. 50% (358 genes) had three or more alleles. To demonstrate the utility of the two reverse genetics platforms, two mutant alleles were isolated for each of the two floral homeotic MADS-box genes, MtPISTILATA and MtAGAMOUS. Molecular and genetic analyses indicate that Tnt1 insertions in exons of both genes are responsible for the defects in floral organ development. In summary, we have developed two efficient reverse genetics platforms to facilitate functional characterization of M. truncatula genes.
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Cheng
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Mingyi
authors.initials: M
authors.lastname: Wang
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Hee-Kyung
authors.initials: HK
authors.lastname: Lee
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Million
authors.initials: M
authors.lastname: Tadege
authors.affiliation: Institut des Sciences du Végétal, CNRS, Avenue de la Terrasse, 91198, Gif sur Yvette Cedex, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Ratet
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Udvardi
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Kirankumar S
authors.initials: KS
authors.lastname: Mysore
authors.affiliation: Division of Plant Biology, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA.
authors.forename: Jiangqi
authors.initials: J
authors.lastname: Wen
mesh_terms: D018744:DNA, Plant / Q000235:genetics; D030561:Databases, Nucleic Acid; D017343:Genes, Plant / Q000235:genetics; D005820:Genetic Testing; D046913:Medicago truncatula / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D012015:Reference Standards; D059386:Reverse Genetics / Q000379:methods*
chemical_list: D018744:DNA, Plant
keywords: Medicago tuncatula; Tnt1 mutant population; database searching; flanking sequence tags (FSTs); floral homeotic genes; reverse genetics