A genome-wide gain-of-function analysis of rice genes using the FOX-hunting system
| Autor: | Mariko Kajikawa, Makoto Hakata, Minami Matsui, Shigeko Ando, Yoshiaki Nagamura, Hirohiko Hirochika, Naoko Toki, Jinhuan Pang, Miki Nakazawa, Akiko Enju, Hidemitsu Nakamura, Takanari Ichikawa, Seiichi Toki, Kazuo Shinozaki, Naokuni Higashi, Kou Amano, Motoaki Seki, Akio Miyao, Hiroaki Ichikawa, Miki Fujita |
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| Rok vydání: | 2007 |
| Předmět: |
Genetics
DNA Complementary Base Sequence Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling Mutant food and beverages Oryza Plant Science General Medicine Biology Genome Phenotype Transformation (genetics) Plasmid Complementary DNA Agronomy and Crop Science Functional genomics Gene Genome Plant DNA Primers Rhizobium |
| Zdroj: | Plant Molecular Biology. 65:357-371 |
| ISSN: | 1573-5028 0167-4412 |
| DOI: | 10.1007/s11103-007-9243-y |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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