Abstract
Forward genetic screens for growth-deficient loss-of-function mutants have uncovered a wide array of genes involved in cell expansion. However, the centrality of cell growth to plant survival means that null mutations in many genes involved in this process are likely to be lethal early in development, making phenotypic analysis difficult. Additionally, the phenotypes of loss-of-function mutations in genes that are members of large gene families might be masked by functional redundancy with other family members. Activation tagging provides a method of screening for dominant overexpression phenotypes in an arbitrarily large collection of transgenic individuals, allowing for functional genomic identification of genes related to cell growth and expansion. In this chapter, we discuss the advantages and limitations of activation tag screening and describe a protocol for identifying activation tag lines with enhanced cell expansion, using dark-grown Arabidopsis thaliana seedlings as an experimental system. We also describe secondary screens to identify candidate genes for further cell biological and genetic characterization. These protocols can be adapted to any process or species of interest, as long as a suitable activation-tagged population and a genome sequence are available.
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References
Cosgrove DJ (2005) Growth of the plant cell wall. Nat Rev Mol Cell Biol 6(11):850–861
Baskin TI (2005) Anisotropic expansion of the plant cell wall. Annu Rev Cell Dev Biol 21:203–222
Dolan L, Davies J (2004) Cell expansion in roots. Curr Opin Plant Biol 7(1):33–39
Weigel D et al (2000) Activation tagging in Arabidopsis. Plant Physiol 122(4):1003–1013
Walden R et al (1994) Activation tagging: a means of isolating genes implicated as playing a role in plant growth and development. Plant Mol Biol 26(5):1521–1528
Jeong DH et al (2002) T-DNA insertional mutagenesis for activation tagging in rice. Plant Physiol 130(4):1636–1644
Wan S et al (2009) Activation tagging, an efficient tool for functional analysis of the rice genome. Plant Mol Biol 69(1–2):69–80
Mur LA et al (2011) Exploiting the Brachypodium Tool Box in cereal and grass research. New Phytol 191(2):334–347
Busov VB et al (2003) Activation tagging of a dominant gibberellin catabolism gene (GA 2-oxidase) from poplar that regulates tree stature. Plant Physiol 132(3):1283–1291
Liu YG, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8(3):457–463
O’Malley RC, Alonso JM, Kim CJ, Leisse TJ, Ecker JR (2007) An adapter ligation-mediated PCR method for high-throughput mapping of T-DNA inserts in the Arabidopsis genome. Nat Protoc 2(11):2910–2917
Gendreau E et al (1997) Cellular basis of hypocotyl growth in Arabidopsis thaliana. Plant Physiol 114(1):295–305
Boron AK, Vissenberg K (2014) The Arabidopsis thaliana hypocotyl, a model to identify and study control mechanisms of cellular expansion. Plant Cell Rep 33:697
Xiao C, Somerville C, Anderson CT (2014) Polygalacturonase involved in cell expansion 1 functions in cell elongation and flower development in Arabidopsis thaliana. Plant Cell 26(3):1018–1035
Kardailsky I et al (1999) Activation tagging of the floral inducer FT. Science 286(5446):1962–1965
van der Graaff E, Dulk-Ras AD, Hooykaas PJ, Keller B (2000) Activation tagging of the LEAFY PETIOLE gene affects leaf petiole development in Arabidopsis thaliana. Development 127(22):4971–4980
van der Graaff E, Hooykaas PJ, Keller B (2002) Activation tagging of the two closely linked genes LEP and VAS independently affects vascular cell number. Plant J 32(5):819–830
Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C (2000) Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. Plant Cell 12(12):2383–2394
Woodward C et al (2005) Interaction of auxin and ERECTA in elaborating Arabidopsis inflorescence architecture revealed by the activation tagging of a new member of the YUCCA family putative flavin monooxygenases. Plant Physiol 139(1):192–203
Xiao C, Chen F, Yu X, Lin C, Fu YF (2009) Over-expression of an AT-hook gene, AHL22, delays flowering and inhibits the elongation of the hypocotyl in Arabidopsis thaliana. Plant Mol Biol 71(1–2):39–50
Mathews H et al (2003) Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. Plant Cell 15(8):1689–1703
Zubko E et al (2002) Activation tagging identifies a gene from Petunia hybrida responsible for the production of active cytokinins in plants. Plant J 29(6):797–808
Marsch-Martinez N (2011) A transposon-based activation tagging system for gene function discovery in Arabidopsis. Methods Mol Biol 754:67–83
Harb A, Pereira A (2013) Activation tagging using the maize En-I transposon system for the identification of abiotic stress resistance genes in Arabidopsis. Methods Mol Biol 1057:193–204
Fladung M, Ahuja MR (1997) Excision of the maize transposable element Ac in periclinal chimeric leaves of 35S-Ac-rolC transgenic aspen-Populus. Plant Mol Biol 33(6):1097–1103
Spena A, Aalen RB, Schulze SC (1989) Cell-autonomous behavior of the rolC gene of Agrobacterium rhizogenes during leaf development: a visual assay for transposon excision in transgenic plants. Plant Cell 1(12):1157–1164
Waki T et al (2013) A GAL4-based targeted activation tagging system in Arabidopsis thaliana. Plant J 73(3):357–367
Sedbrook JC, Ehrhardt DW, Fisher SE, Scheible WR, Somerville CR (2004) The Arabidopsis sku6/spiral1 gene encodes a plus end-localized microtubule-interacting protein involved in directional cell expansion. Plant Cell 16(6):1506–1520
Acknowledgments
Thanks to Chris Somerville and members of the Somerville lab for helpful discussions on this topic, to Wenting Xi for technical assistance with primary and secondary screening, and to William Barnes for critical reading. The writing of this chapter was supported as part of the Center for Lignocellulose Structure and Formation, an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Basic Energy Sciences under Award # DE-SC0001090.
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Xiao, C., Anderson, C.T. (2015). Activation Tag Screening for Cell Expansion Genes in Arabidopsis thaliana . In: Estevez, J. (eds) Plant Cell Expansion. Methods in Molecular Biology, vol 1242. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1902-4_14
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DOI: https://doi.org/10.1007/978-1-4939-1902-4_14
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