Abstract
Although well-accepted as the ultimate method for cotton functional genomics, Agrobacterium tumefaciens-mediated cotton transformation is not widely used for functional analyses of cotton genes and their promoters since regeneration of cotton in tissue culture is lengthy and labor intensive. In certain cases, A. rhizogenes-induced hairy root culture has been a suitable molecular tool for functional analyses of genes and promoters for plants that are difficult to regenerate by A. tumefaciens-mediated transformation. Similarly, A. rhizogenes-induced hairy root cultures are an alternative tool for cotton functional genomics. In this chapter, the advantages and disadvantages of using A. rhizogenes-induced cotton hairy root culture over A. tumefaciens-mediated cotton transformation are discussed. The procedures for transformation, generation, selection, and molecular analyses of transgenic cotton hairy roots are introduced by describing the functional analysis of a cotton promoter in cotton hairy roots generated by A. rhizogenes-mediated transformation.
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References
Veena V, Taylor CG (2007) Agrobacterium rhizogenes: recent developments and promising applications. In Vitro Cell Dev Biol Plant 43:383–403
Wang S, Wang JW, Yu N, Li CH, Luo B, Gou JY, Wang LJ, Chen XY (2004) Control of plant trichome development by a cotton fiber MYB gene. Plant Cell 16:2323–2334
Delaney SK, Orford SJ, Martin-Harris M, Timmis JN (2007) The fiber specificity of the cotton FSltp4 gene promoter is regulated by an AT-rich promoter region and the AT-Hook transcription factor GhAT1. Plant Cell Physiol 48:1426–1437
Guan X, Lee JJ, Pang M, Shi X, Stelly DM, Chen ZJ (2011) Activation of Arabidopsis seed hair development by cotton fiber-related genes. PLoS One 6:e21301
Limpens E, Ramos J, Franken C, Raz V, Compaan B, Franssen H, Bisseling T, Geurts R (2004) RNA interference in Agrobacterium rhizogenes-transformed roots of Arabidopsis and Medicago truncatula. J Exp Bot 55:983–992
Nontachaiyapoom S, Scott PT, Men AE, Kinkema M, Schenk PM, Gresshoff PM (2006) Promoters of orthologous Glycine max and Lotus japonicus nodulation autoregulation genes interchangeably drive phloem-specific expression in transgenic plants. Mol Plant Microbe Interact 20:769–780
Moreno-Valenzuela OA, Minero-Garcia Y, Brito-Argaez L, Carbajal-Mora E, Echeverria O, Vazquez-Nin G et al (2003) Immunocytolocalization of tryptophan decarboxylase in Catharanthus roseus hairy roots. Mol Biotechnol 23:11–18
Marjamaa K, Hilden K, Kukkola E, Lehtonen M, Holkeri H, Haapaniemi P et al (2006) Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies). Plant Mol Biol 61:719–732
Grønlund M, Roussis A, Flemetakis E, Quaedvlieg NEM, Schlaman HRM, Umehara Y et al (2005) Analysis of promoter activity of the early nodulin Enod40 in Lotus japonicas. Mol Plant Microbe Interact 18:414–427
Triplett BA, Moss SC, Bland JM, Dowd MK (2008) Induction of hairy root cultures from Gossypium hirsutum and Gossypium barbadense to produce gossypol and related compounds. In Vitro Cell Dev Biol Plant 44:508–517
Frankfater CR, Dowd MK, Triplett BA (2009) Effect of elicitors on the production of gossypol and methylated gossypol in cotton hairy roots. Plant Cell Tiss Org Cult 98:341–349
Verma PC, Trivedi I, Singh H, Shukla AK, Kumar M, Upadhyay SK et al (2009) Efficient production of gossypol from hairy root cultures of cotton (Gossypium hirsutum L.). Curr Pharm Biotechnol 10:691–700
Wubben M, Callahan FE, Triplett BA, Jenkins JN (2009) Phenotypic and molecular evaluation of cotton hairy roots as a model system for studying nematode interactions. Plant Cell Rep 28:1399–1409
Kim HJ, Murai N, Fang DD, Triplett BA (2011) Functional analysis of Gossypium hirsutum cellulose synthase catalytic subunit 4 promoter in transgenic Arabidopsis and cotton tissues. Plant Sci 180:323–332
Trolinder N (2010) Cotton regeneration. In: Stewart JM, Oosterhuis DM, Heitholt JJ, Mauney JR (eds) Physiology of cotton. Springer, New York, pp 379–382
Chabaud M, Boisson-Dernier A, Zhang J, Taylor CG, Yu O, Barker DG (2006) Agrobacterium rhizogenes-mediated root transformation. In: Mathesius U, Journet EP, Sumner LW (eds) Medicago truncatula handbook. ISBN 0-9754303-1-9. http://www.noble.org/MedicagoHandbook/
Roberts CS, Rajagopal S, Smith LA, Nguyen TA, Yang W, Nugroho S, Ravi KS, Cao M, Vijhayachandra K, Patell V, Harcourt RL, Dransfield L, Desamero N, Slamet I, Keese P, Kilian A, Jefferson RA (1998) A comprehensive set of modular vectors for advanced manipulations and efficient transformation of plants by both Agrobacterium and direct DNA uptake methods. pCAMBIA vector release manual version 3.05
Jefferson RA, Kayanagh TA, Bevan MW (1987) GUS fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Höfgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res 16:9877
Nagel R, Elliott A, Masel A, Birch RG, Manners JM (1990) Electroporation of binary Ti plasmid vector into Agrobacterium tumefaciens and Agrobacterium rhizogenes. FEMS Microbiol Lett 67:325–328
Nilsson O, Olsson O (1997) Getting to the root: the role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots. Physiol Plant 100:463–473
Acknowledgments
This work was supported by United States Department of Agriculture-Agricultural Research Service project 6435-21000-016-00D. The author acknowledges Stephanie Moss, Cheryl Frankfater, Barbara Triplett, and Michael Dowd of USDA-ARS-Southern Regional Research Center who initiated cotton hairy root cultures to produce gossypol compounds. The author also thanks Dr. Barbara Triplett for critically reading manuscript.
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Kim, H.J. (2013). Agrobacterium rhizogenes-Induced Cotton Hairy Root Culture as an Alternative Tool for Cotton Functional Genomics. In: Zhang, B. (eds) Transgenic Cotton. Methods in Molecular Biology, vol 958. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-212-4_15
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DOI: https://doi.org/10.1007/978-1-62703-212-4_15
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