title: Tomato TILLING technology: development of a reverse genetics tool for the efficient isolation of mutants from Micro-Tom mutant libraries.
abstract: To accelerate functional genomic research in tomato, we developed a Micro-Tom TILLING (Targeting Induced Local Lesions In Genomes) platform. DNA pools were constructed from 3,052 ethyl methanesulfonate (EMS) mutant lines treated with 0.5 or 1.0% EMS. The mutation frequency was calculated by screening 10 genes. The 0.5% EMS population had a mild mutation frequency of one mutation per 1,710 kb, whereas the 1.0% EMS population had a frequency of one mutation per 737 kb, a frequency suitable for producing an allelic series of mutations in the target genes. The overall mutation frequency was one mutation per 1,237 kb, which affected an average of three alleles per kilobase screened. To assess whether a Micro-Tom TILLING platform could be used for efficient mutant isolation, six ethylene receptor genes in tomato (SlETR1-SlETR6) were screened. Two allelic mutants of SlETR1 (Sletr1-1 and Sletr1-2) that resulted in reduced ethylene responses were identified, indicating that our Micro-Tom TILLING platform provides a powerful tool for the rapid detection of mutations in an EMS mutant library. This work provides a practical and publicly accessible tool for the study of fruit biology and for obtaining novel genetic material that can be used to improve important agronomic traits in tomato.
authors.affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Okabe
authors.forename: Erika
authors.initials: E
authors.lastname: Asamizu
authors.forename: Takeshi
authors.initials: T
authors.lastname: Saito
authors.forename: Chiaki
authors.initials: C
authors.lastname: Matsukura
authors.forename: Tohru
authors.initials: T
authors.lastname: Ariizumi
authors.forename: Cécile
authors.initials: C
authors.lastname: Brès
authors.forename: Christophe
authors.initials: C
authors.lastname: Rothan
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Mizoguchi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ezura
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D018744:DNA, Plant / Q000235:genetics; D005020:Ethyl Methanesulfonate; D005030:Ethylenes / Q000378:metabolism; D005638:Fruit / Q000502:physiology; D015723:Gene Library; D017343:Genes, Plant; D023281:Genomics / Q000379:methods*; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D059645:Mutation Rate; D010940:Plant Proteins / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000235:genetics*
chemical_list: D018744:DNA, Plant; D005030:Ethylenes; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C083750:ethylene receptors, plant; C036216:ethylene; D005020:Ethyl Methanesulfonate