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      The maize ( Zea mays L.) roothairless3 gene encodes a putative GPI-anchored, monocot-specific, COBRA-like protein that significantly affects grain yield

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          Abstract

          Summary

          The rth3 ( roothairless 3) mutant is specifically affected in root hair elongation. We report here the cloning of the rth3 gene via a PCR-based strategy (amplification of insertion mutagenized sites) and demonstrate that it encodes a COBRA-like protein that displays all the structural features of a glycosylphosphatidylinositol anchor. Genes of the COBRA family are involved in various types of cell expansion and cell wall biosynthesis. The rth3 gene belongs to a monocot-specific clade of the COBRA gene family comprising two maize and two rice genes. While the rice ( Oryza sativa) gene OsBC1L1 appears to be orthologous to rth3 based on sequence similarity (86% identity at the protein level) and maize/rice synteny, the maize ( Zea mays L.) rth3-like gene does not appear to be a functional homolog of rth3 based on their distinct expression profiles. Massively parallel signature sequencing analysis detected rth3 expression in all analyzed tissues, but at relatively low levels, with the most abundant expression in primary roots where the root hair phenotype is manifested. In situ hybridization experiments confine rth3 expression to root hair-forming epidermal cells and lateral root primordia. Remarkably, in replicated field trials involving near-isogenic lines, the rth3 mutant conferred significant losses in grain yield.

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          Most cited references50

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          Protein Identification and Analysis Tools on the ExPASy Server

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            Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays.

            We describe a novel sequencing approach that combines non-gel-based signature sequencing with in vitro cloning of millions of templates on separate 5 microm diameter microbeads. After constructing a microbead library of DNA templates by in vitro cloning, we assembled a planar array of a million template-containing microbeads in a flow cell at a density greater than 3x10(6) microbeads/cm2. Sequences of the free ends of the cloned templates on each microbead were then simultaneously analyzed using a fluorescence-based signature sequencing method that does not require DNA fragment separation. Signature sequences of 16-20 bases were obtained by repeated cycles of enzymatic cleavage with a type IIs restriction endonuclease, adaptor ligation, and sequence interrogation by encoded hybridization probes. The approach was validated by sequencing over 269,000 signatures from two cDNA libraries constructed from a fully sequenced strain of Saccharomyces cerevisiae, and by measuring gene expression levels in the human cell line THP-1. The approach provides an unprecedented depth of analysis permitting application of powerful statistical techniques for discovery of functional relationships among genes, whether known or unknown beforehand, or whether expressed at high or very low levels.
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              Identification of novel genes in Arabidopsis involved in secondary cell wall formation using expression profiling and reverse genetics.

              Forward genetic screens have led to the isolation of several genes involved in secondary cell wall formation. A variety of evidence, however, suggests that the list of genes identified is not exhaustive. To address this problem, microarray data have been generated from tissue undergoing secondary cell wall formation and used to identify genes that exhibit a similar expression pattern to the secondary cell wall-specific cellulose synthase genes IRREGULAR XYLEM1 (IRX1) and IRX3. Cross-referencing this analysis with publicly available microarray data resulted in the selection of 16 genes for reverse genetic analysis. Lines containing an insertion in seven of these genes exhibited a clear irx phenotype characteristic of a secondary cell wall defect. Only one line, containing an insertion in a member of the COBRA gene family, exhibited a large decrease in cellulose content. Five of the genes identified as being essential for secondary cell wall biosynthesis have not been previously characterized. These genes are likely to define entirely novel processes in secondary cell wall formation and illustrate the success of combining expression data with reverse genetics to address gene function.
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                Author and article information

                Journal
                Plant J
                tpj
                The Plant Journal
                Blackwell Publishing Ltd
                0960-7412
                1365-313X
                June 2008
                14 April 2008
                : 54
                : 5
                : 888-898
                Affiliations
                [1 ]Center for Plant Molecular Biology, Department of General Genetics, Eberhard-Karls-University Tuebingen 72076 Tuebingen, Germany
                [2 ]Department of Agronomy, Iowa State University Ames, IA 50011, USA
                [3 ]Institute for General Botany and Botanical Garden, University of Hamburg 22609 Hamburg, Germany
                [4 ]Pioneer Hi-Bred International, Inc. – a DuPont Company Johnston, IA 50131, USA
                [5 ]Department of Genetics, Development, and Cell Biology, Iowa State University Ames, IA 50011, USA
                [6 ]Center for Plant Genomics, Iowa State University , Ames, IA 50011-36506, USA
                Author notes
                *For correspondence (fax +1 515 294 5256; e-mail schnable@ 123456iastate.edu ).
                [†]

                Present address: 6416 E. Lake, Sammamish Parkway NE, Redmond, WA 98052, USA.

                [‡]

                Present address: BASF Future Business GmbH, 67063 Ludwigshafen, Germany.

                [§]

                Present address: BASF Plant Science, Research Triangle Park, NC 27709, USA.

                [¶]

                These authors contributed equally to this paper.

                Article
                10.1111/j.1365-313X.2008.03459.x
                2440564
                18298667
                73a14f09-7be2-4e46-9479-292150abf3d6
                © 2008 The Authors Journal compilation © 2008 Blackwell Publishing Ltd

                Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

                History
                : 13 November 2007
                : 24 January 2008
                : 07 February 2008
                Categories
                Original Articles

                Plant science & Botany
                root hairs,maize,mutant,rth3,cobra-like
                Plant science & Botany
                root hairs, maize, mutant, rth3, cobra-like

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