title: A rice gene activation/knockout mutant resource for high throughput functional genomics.
abstract: Using transfer DNA (T-DNA) with functions of gene trap and gene knockout and activation tagging, a mutant population containing 55,000 lines was generated. Approximately 81% of this population carries 1-2 T-DNA copies per line, and the retrotransposon Tos17 was mostly inactive in this population during tissue culture. A total of 11,992 flanking sequence tags (FSTs) have been obtained and assigned to the rice genome. T-DNA was preferentially ( approximately 80%) integrated into genic regions. A total of 19,000 FSTs pooled from this and another T-DNA tagged population were analyzed and compared with 18,000 FSTs from a Tos17 tagged population. There was difference in preference for integrations into genic, coding, and flanking regions, as well as repetitive sequences and centromeric regions, between T-DNA and Tos17; however, T-DNA integration was more evenly distributed in the rice genome than Tos17. Our T-DNA contains an enhancer octamer next to the left border, expression of genes within genetics distances of 12.5 kb was enhanced. For example, the normal height of a severe dwarf mutant, with its gibberellin 2-oxidase (GA2ox) gene being activated by T-DNA, was restored upon GA treatment, indicating GA2ox was one of the key enzymes regulating the endogenous level of GA. Our T-DNA also contains a promoterless GUS gene next to the right border. GUS activity screening facilitated identification of genes responsive to various stresses and those regulated temporally and spatially in large scale with high frequency. Our mutant population offers a highly valuable resource for high throughput rice functional analyses using both forward and reverse genetic approaches.
authors.affiliation: Institute of Plant and Microbial Biology, Academia Sinica, Nankang, Taipei 115, Taiwan, ROC.
authors.forename: Yue-Ie
authors.initials: YI
authors.lastname: Hsing
authors.forename: Chyr-Guan
authors.initials: CG
authors.lastname: Chern
authors.forename: Ming-Jen
authors.initials: MJ
authors.lastname: Fan
authors.forename: Po-Chang
authors.initials: PC
authors.lastname: Lu
authors.forename: Ku-Ting
authors.initials: KT
authors.lastname: Chen
authors.forename: Shuen-Fang
authors.initials: SF
authors.lastname: Lo
authors.forename: Peng-Kai
authors.initials: PK
authors.lastname: Sun
authors.forename: Shin-Lon
authors.initials: SL
authors.lastname: Ho
authors.forename: Kuo-Wei
authors.initials: KW
authors.lastname: Lee
authors.forename: Yi-Chieh
authors.initials: YC
authors.lastname: Wang
authors.forename: Wen-Lii
authors.initials: WL
authors.lastname: Huang
authors.forename: Swee-Suak
authors.initials: SS
authors.lastname: Ko
authors.forename: Shu
authors.initials: S
authors.lastname: Chen
authors.forename: Jyh-Long
authors.initials: JL
authors.lastname: Chen
authors.forename: Chun-I
authors.initials: CI
authors.lastname: Chung
authors.forename: Yao-Cheng
authors.initials: YC
authors.lastname: Lin
authors.forename: Ai-Ling
authors.initials: AL
authors.lastname: Hour
authors.forename: Yet-Walt
authors.initials: YW
authors.lastname: Wang
authors.forename: Ya-Chi
authors.initials: YC
authors.lastname: Chang
authors.forename: Min-Wei
authors.initials: MW
authors.lastname: Tsai
authors.forename: Yi-Show
authors.initials: YS
authors.lastname: Lin
authors.forename: Yin-Chin
authors.initials: YC
authors.lastname: Chen
authors.forename: Hsing-Mu
authors.initials: HM
authors.lastname: Yen
authors.forename: Charng-Pei
authors.initials: CP
authors.lastname: Li
authors.forename: Chiu-Kai
authors.initials: CK
authors.lastname: Wey
authors.forename: Ching-Shan
authors.initials: CS
authors.lastname: Tseng
authors.forename: Ming-Hsing
authors.initials: MH
authors.lastname: Lai
authors.forename: Sheng-Chung
authors.initials: SC
authors.lastname: Huang
authors.forename: Liang-Jwu
authors.initials: LJ
authors.lastname: Chen
authors.forename: Su-May
authors.initials: SM
authors.lastname: Yu
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005822:Genetic Vectors; D023281:Genomics; D009154:Mutation; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified; D016324:Sequence Tagged Sites; D015533:Transcriptional Activation
chemical_list: D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA