Abstract
This chapter describes an efficient Agrobacterium-mediated genetic transformation of lentil by use of cotyledonary node explants, an optimized wounding method, and vacuum infiltration. Transformation protocol was followed by direct regeneration of transgenic shoots and micrografting of the shoots on root stocks to obtain whole-plant regeneration. The most efficient transgene expression on the axil region was obtained when the Agrobacterium KYRT1 strain was used. Gradually increasing selection pressure and repeated removal of regenerated shoots between selection steps increased the number of transgene-expressing shoots greatly. This protocol allowed 2.3 % transformation efficiency and stable transgene expression and transmission which were tracked through three generations.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Zohary D, Hopf M (1988) Domestication of plants in the old world. Clarendon, London, UK
Muehlbauer FJ, Kaiser WJ, Clement SL, Summerfield RJ (1995) Production and breeding of lentil. In: Sparks D (ed) Advances in agronomy. Academic, New York, NY, pp 283–332
Murphy DJ (2007) People, plants and genes. Oxford University Press, New York, NY
Chowrira GM, Cavileer TD, Gupta SK, Lurquin PF, Berger PH (1998) Coat protein-mediated resistance to pea enation mosaic virus in transgenic Pisum sativum L. Transgenic Res 7:265–271
Chowrira GM (1996) Transgenic grain legumes obtained by in planta electroporation-mediated gene transfer. Mol Biotechnol 5: 85–95
Sommers DA, Samac DA, Olhoft PM (2003) Recent advances in legume transformation. Plant Physiol 131:892–899
Gulati A, Schryer P, McHughen A (2002) Production of fertile transgenic lentil (Lens culinaris Medik) plants using particle bombardment. In Vitro Cell Dev-Pl 38:316–324
Chopra R, Raman SA (2012) Use of sonication and vacuum infiltration for Agrobacterium – mediated transformation of an Indian lentil (Lens culinaris Medik.) cultivar. Sci Hortic 143:127–134
Khatib F, Makris A, Yamaguchi-Shinozaki K, Kumar S, Sarker A, Erskine W, Baum M (2011) Expression of the DREB1A gene in lentil (Lens culinaris Medik. subsp culinaris) transformed with the Agrobacterium system. Crop Pasture Sci 62:488–495
Torisky RS, Kovacs L, Avdiushko S, Newman JD, Hunt AG, Collins GB (1997) Development of a binary vector system for plant transformation based on the supervirulent Agrobacterium tumefaciens strain Chry5. Plant Cell Rep 17:102–108
Kapila J, De Rycke R, Van Montagu M, Angenon G (1997) An Agrobacterium-mediated transient gene expression system for intact leaves. Plant Sci 122:101–108
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Christou P (1993) Control of plant gene expression. CRC Press, Boca Raton, FL
Hawtin GC (1980) Advances in legume science. In: Muehlbauer FJ (ed) Advances in agronomy. Academic, New York, NY, pp 283–332
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Akcay, U.C., Yücel, M., Öktem, H.A. (2015). Lentil (Lens culinaris Medik). In: Wang, K. (eds) Agrobacterium Protocols. Methods in Molecular Biology, vol 1223. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-1695-5_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1695-5_21
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4939-1694-8
Online ISBN: 978-1-4939-1695-5
eBook Packages: Springer Protocols