title: A genome-wide gain-of function analysis of rice genes using the FOX-hunting system.
abstract: The latest report has estimated the number of rice genes to be approximately 32,000. To elucidate the functions of a large population of rice genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the Full-length cDNA Over-eXpresser (FOX) gene-hunting system. This system is very useful for analyzing various gain-of-function phenotypes from large populations of transgenic plants overexpressing cDNAs of interest and others with unknown or important functions. We collected the plasmid DNAs of 13,980 independent full-length cDNA (FL-cDNA) clones to produce a FOX library by placing individual cDNAs under the control of the maize Ubiquitin-1 promoter. The FOX library was transformed into rice by Agrobacterium-mediated high-speed transformation. So far, we have generated approximately 12,000 FOX-rice lines. Genomic PCR analysis indicated that the average number of FL-cDNAs introduced into individual lines was 1.04. Sequencing analysis of the PCR fragments carrying FL-cDNAs from 8615 FOX-rice lines identified FL-cDNAs in 8225 lines, and a database search classified the cDNAs into 5462 independent ones. Approximately 16.6% of FOX-rice lines examined showed altered growth or morphological characteristics. Three super-dwarf mutants overexpressed a novel gibberellin 2-oxidase gene,confirming the importance of this system. We also show here the other morphological alterations caused by individual FL-cDNA expression. These dominant phenotypes should be valuable indicators for gene discovery and functional analysis.
authors.affiliation: Division of Genome and Biodiversity Research, NationalInstitute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.
authors.forename: Hidemitsu
authors.initials: H
authors.lastname: Nakamura
authors.forename: Makoto
authors.initials: M
authors.lastname: Hakata
authors.forename: Kou
authors.initials: K
authors.lastname: Amano
authors.forename: Akio
authors.initials: A
authors.lastname: Miyao
authors.forename: Naoko
authors.initials: N
authors.lastname: Toki
authors.forename: Mariko
authors.initials: M
authors.lastname: Kajikawa
authors.forename: Jinhuan
authors.initials: J
authors.lastname: Pang
authors.forename: Naokuni
authors.initials: N
authors.lastname: Higashi
authors.forename: Shigeko
authors.initials: S
authors.lastname: Ando
authors.forename: Seiichi
authors.initials: S
authors.lastname: Toki
authors.forename: Miki
authors.initials: M
authors.lastname: Fujita
authors.forename: Akiko
authors.initials: A
authors.lastname: Enju
authors.forename: Motoaki
authors.initials: M
authors.lastname: Seki
authors.forename: Miki
authors.initials: M
authors.lastname: Nakazawa
authors.forename: Takanari
authors.initials: T
authors.lastname: Ichikawa
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shinozaki
authors.forename: Minami
authors.initials: M
authors.lastname: Matsui
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Nagamura
authors.forename: Hirohiko
authors.initials: H
authors.lastname: Hirochika
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ichikawa
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary; D020869:Gene Expression Profiling / Q000379:methods*; D018745:Genome, Plant; D012275:Oryza / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary