TY - JOUR A1 - Hilson, Pierre A1 - Allemeersch, Joke A1 - Altmann, Thomas A1 - Aubourg, Sebastien A1 - Avon, Alexandra A1 - Beynon, Jim A1 - Bhalerao, Rishikesh P. A1 - Bitton, Frederique A1 - Caboche, Michel A1 - Cannoot, Bernard A1 - Chardakov, Vasil A1 - Cognet-Holliger, Cecile A1 - Colot, Vincent A1 - Crowe, Mark A1 - Darimont, Caroline A1 - Durinck, Steffen A1 - Eickhoff, Holger A1 - deLongevialle, Andeol Falcon A1 - Farmer, Edward E. A1 - Grant, Murray A1 - Kuiper, Martin T. R. A1 - Lehrach, Hans A1 - Leon, Celine A1 - Leyva, Antonio A1 - Lundeberg, Joakim A1 - Lurin, Claire A1 - Moreau, Yves T1 - Versatile gene-specific sequence tags for arabidopsis functional genomics : transcript profiling and reserve genetics applications N2 - Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics Y1 - 2004 ER - TY - JOUR A1 - Robert, Helene S. A1 - Grunewald, Wim A1 - Sauer, Michael A1 - Cannoot, Bernard A1 - Soriano, Mercedes A1 - Swarup, Ranjan A1 - Weijers, Dolf A1 - Bennett, Malcolm A1 - Boutilier, Kim A1 - Friml, Jiri T1 - Plant embryogenesis requires AUX/LAX-mediated auxin influx JF - Development : Company of Biologists N2 - The plant hormone auxin and its directional transport are known to play a crucial role in defining the embryonic axis and subsequent development of the body plan. Although the role of PIN auxin efflux transporters has been clearly assigned during embryonic shoot and root specification, the role of the auxin influx carriers AUX1 and LIKE-AUX1 (LAX) proteins is not well established. Here, we used chemical and genetic tools on Brassica napus microspore-derived embryos and Arabidopsis thaliana zygotic embryos, and demonstrate that AUX1, LAX1 and LAX2 are required for both shoot and root pole formation, in concert with PIN efflux carriers. Furthermore, we uncovered a positive-feedback loop between MONOPTEROS-(ARF5)dependent auxin signalling and auxin transport. This MONOPTEROS dependent transcriptional regulation of auxin influx (AUX1, LAX1 and LAX2) and auxin efflux (PIN1 and PIN4) carriers by MONOPTEROS helps to maintain proper auxin transport to the root tip. These results indicate that auxin-dependent cell specification during embryo development requires balanced auxin transport involving both influx and efflux mechanisms, and that this transport is maintained by a positive transcriptional feedback on auxin signalling. KW - Arabidopsis thaliana embryogenesis KW - Auxin transport KW - AUX1 KW - LIKE-AUX1 (LAX) KW - MONOPTEROS (ARF5) KW - PIN KW - Brassica napus KW - Microspore Y1 - 2015 U6 - https://doi.org/10.1242/dev.115832 SN - 0950-1991 SN - 1477-9129 VL - 142 IS - 4 SP - 702 EP - 711 PB - Company of Biologists Limited CY - Cambridge ER -