Abstract
A large population of Medicago truncatula insertion lines has been generated using the Tnt1 retrotransposon. More than 21,000 insertion lines have been generated, representing more than 500,000 insertion events. This mutant population is being used by the legume research community to screen for various different mutants using a forward genetics approach. Some of the phenotypes that have been screened using this population include developmentally abnormal phenotypes in leaves, stem, flowers, and roots. In addition to these, mutants with defects in symbiosis with Rhizobium and mycorrhiza, mutants with altered nonhost resistance against Asian Soybean Rust and switch grass rust pathogens, mutants with altered lignin content, mutants with altered cell wall structure, etc. have been identified. Here, we describe the high throughput methodology that is being used to identify these M. truncatula mutants.
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
Graham PH, Vance CP (2003) Legumes: importance and constraints to greater use. Plant Physiol 131:872–877
Tadege M et al (2008) Large-scale insertional mutagenesis using the Tnt1 retrotransposon in the model legume Medicago truncatula. Plant J 54:335–347
Kumar A, Bennetzen JL (1999) Plant retrotransposons. Ann Rev Genet 33:479–532
Perez-Hormaeche J et al (2008) Invasion of the Arabidopsis genome by the tobacco retrotransposon Tnt1 is controlled by reversible transcriptional gene silencing. Plant Physiol 147:1264–1278
d’Erfurth I et al (2003) Efficient transposition of the Tnt1 tobacco retrotransposon in the model legume Medicago truncatula. Plant J 34:95–106
Grandbastien MA (1997) The expression of the tobacco Tnt1 retrotransposon is linked to plant defense responses. Genetica 100:241–252
Greco R et al (2001) Transposon insertional mutagenesis in rice. Plant Physiol 125:1175–1177
Mazier M et al (2007) Successful gene tagging in lettuce using the Tnt1 retrotransposon from tobacco. Plant Physiol 144:18–31
Cui Y et al (2013) Tnt1 retrotransposon mutagenesis: a tool for soybean functional genomics. Plant Physiol 161:36–47
Tadege M et al (2009) Mutagenesis and beyond! Tools for understanding legume biology. Plant Physiol 151:978–984
Cheng X et al (2011) Reverse genetics in Medicago truncatula using Tnt1 insertion mutants. Meth Mol Biol 678:179–190
Pislariu CI et al (2012) A Medicago truncatula tobacco retrotransposon insertion mutant collection with defects in nodule development and symbiotic nitrogen fixation. Plant Physiol 159:1686–1699
Mulligan JT, Long SR (1985) Induction of Rhizobium meliloti nodC expression by plant exudate requires nodD. Proc Nat Acad Sci USA 82:6609–6613
Ratet P et al (2010) Tnt1 induced mutations in Medicago: characterization and applications. In: Meksem K, Kahl G (eds) The handbook of plant mutation screening: mining of natural and induced alleles. Wiley-Blackwell, Chichester, pp 83–99
Acknowledgments
This work was supported by the Samuel Roberts Noble Foundation and NSF Plant Genome Program Grants (DBI-0703285 and IOS-1127155).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Yarce, J.C.S., Lee, HK., Tadege, M., Ratet, P., Mysore, K.S. (2013). Forward Genetics Screening of Medicago truncatula Tnt1 Insertion Lines. In: Rose, R. (eds) Legume Genomics. Methods in Molecular Biology, vol 1069. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-613-9_8
Download citation
DOI: https://doi.org/10.1007/978-1-62703-613-9_8
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-612-2
Online ISBN: 978-1-62703-613-9
eBook Packages: Springer Protocols