Establishment of a patterned GAL4-VP16 transactivation system for discovering gene function in rice

• Published in: The Plant journal : for cell and molecular biology Vol. 46; no. 6; pp. 1059 - 1072
• Main Authors: Liang, Dacheng, Wu, Changyin, Li, Caishun, Xu, Caiguo, Zhang, Jianwei, Kilian, Andrzej, Li, Xianghua, Zhang, Qifa, Xiong, Lizhong
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.06.2006; Blackwell Publishing Ltd; Blackwell Science
• Subjects: Biological and medical sciences; cultivars; DNA-Binding Proteins; ectopic expression; flowering; Flowers - genetics; Flowers - metabolism; Fundamental and applied biological sciences. Psychology; GAL4–UAS; Gene Expression Profiling; Gene Expression Regulation, Plant; genes; Genes, Plant - genetics; growth retardation; Herpes Simplex Virus Protein Vmw65 - genetics; Herpes Simplex Virus Protein Vmw65 - metabolism; hybrids; leaves; Molecular and cellular biology; Molecular genetics; Oryza - genetics; Oryza - metabolism; Oryza sativa; pattern line; Phenotype; pistil; Plant Leaves - genetics; Plant Leaves - metabolism; Plant Proteins - genetics; Plant Proteins - metabolism; Plants, Genetically Modified; rice; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Seedlings - genetics; Seedlings - metabolism; stigma; target line; tillers; transcription (genetics); transcription factor; transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription. Transcription factor. Splicing. Rna processing; Transcriptional Activation; Anther; Monocotyledones; ectopic expression; Growth; target line; Plant leaf; Oryza sativa; Gene; Gramineae; pattern line; Angiospermae; GAL4-UAS; Flowering; Spermatophyta; Transcription factor
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/j.1365-313x.2006.02747.x

A binary GAL4-VP16-UAS transactivation system has been established in rice (Oryza sativa L.) in this study for the discovery of gene functions. This binary system consists of two types of transgenic lines, pattern lines and target lines. The pattern lines were produced by transformation of Zhonghua 11, a japonica cultivar, with a construct consisting of the transactivator gene GAL4-VP16 controlled by a minimal promoter and the GUSplus reporter controlled by the upstream activation sequence (UAS; cis-element to GAL4). Target lines were generated by transformation of Zhonghua 11 with constructs carrying the EGFP reporter and target genes of interest, both controlled by the UAS but in opposite directions. Hybrid plants were obtained by crossing target lines of 10 putative transcription factor genes from rice with six pattern lines showing expression in anther, stigma, palea, lemma and leaves. The EGFP and target genes perfectly co-expressed in hybrid plants with the same expression patterns as in the pattern lines. Various phenotypic changes, such as delayed flowering, multiple pistils, dwarfism, narrow and droopy leaves, reduced tillers, growth retardation and sterility, were induced as a result of the expression of the target genes. It is concluded that this transactivation system can provide a useful tool in rice to unveil latent functions of unknown or known genes.