Establishment of a Lotus japonicus gene tagging population using the exon-targeting endogenous retrotransposon LORE1.
We established a gene tagging population of the model legume Lotus japonicus using an endogenous long terminal repeat (LTR) retrotransposon Lotus Retrotransposon 1 (LORE1). The population was composed of 2450 plant lines, from which a total of 4532 flanking sequence tags of LORE1 were recovered by pyrosequencing. The two-dimensional arrangement of the plant population, together with the use of multiple identifier sequences in the primers used to amplify the flanking regions, made it possible to trace insertions back to the original plant lines. The large-scale detection of new LORE1 insertion sites revealed a preference for genic regions, especially in exons of protein-coding genes, which is an interesting feature to consider in the interaction between host genomes and chromoviruses, to which LORE1 belongs, a class of retrotransposon widely distributed among plants. Forward screening of the symbiotic mutants from the population succeeded to identify five symbiotic mutants of known genes. These data suggest that LORE1 is robust as a genetic tool.
Division of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan.
Eigo
E
Fukai
Takashi
T
Soyano
Yosuke
Y
Umehara
Shinobu
S
Nakayama
Hideki
H
Hirakawa
Satoshi
S
Tabata
Shusei
S
Sato
Makoto
M
Hayashi
D017931:DNA Primers / Q000235:genetics; D005091:Exons / Q000235:genetics*; D018390:Gene Targeting; D000070116:Lotus / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D009154:Mutation; D018626:Retroelements / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013559:Symbiosis; D020079:Terminal Repeat Sequences / Q000235:genetics
D017931:DNA Primers; D018626:Retroelements