Abstract
The combination of the DNA sequence-specific recombination system Cre/LoxP and the DNA transposon system Activator (Ac)/Dissociation (Ds) has been used for insertional and deletional mutagenesis, as well as for the generation of artificial ring chromosomes in model plants such as Arabidopsis and tobacco. However, it takes a long time to complete this process, even in Arabidopsis. To overcome this issue, a new binary vector, pDLHC, has been developed to induce chromosomal rearrangements for a short time in rice. pDLHC has been found to be effective in the induction of deletions between two LoxPs in the T2 generation of “Nihon bare” expressing Ac TPase. pDLHC has potential for the efficient generation of various types of chromosomal rearrangements including deletions, inversions, translocations and artificial ring chromosomes in plants, and the detailed protocol for rice is described here.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Medberry SL, Dale E, Qin M, Ow DW (1995) Intra-chromosomal rearrangements generated by Cre-lox site-specific recombination. Nucleic Acids Res 23:485–490
Woody ST, Austin-Phillips S, Amasino RM, Krysan PJ (2007) The WiscDsLox T-DNA collection: an arabidopsis community resource generated by using an improved high-throughput T-DNA sequencing pipeline. J Plant Res 120:157–165
Zhang S, Raina S, Li H, Li J, Dec E, Ma H, Huang H, Fedoroff NV (2003) Resources for targeted insertional and deletional mutagenesis in Arabidopsis. Plant Mol Biol 53:133–150
Murata M, Shibata F, Hironaka A, Kashihara K, Fujimoto S, Yokota E, Nagaki K (2013) Generation of an artificial ring chromosome in Arabidopsis by Cre/LoxP-mediated recombination. Plant J 74:363–371
Ito Y, Eiguchi M, Kurata N (2004) Establishment of an enhancer trap system with Ds and GUS for functional genomics in rice. Mol Genet Genomics 271:639–650
Ohta S, Mita S, Hattori T, Nakamura K (1990) Construction and expression in tobacco of a beta-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31:805–813
Okuzaki A, Shimizu T, Kaku K, Kawai K, Toriyama K (2007) A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice. Plant Mol Biol 64:219–224
Cuellar W, Gaudin A, Solorzano D, Casas A, Nopo L, Chudalayandi P, Medrano G, Kreuze J, Ghislain M (2006) Self-excision of the antibiotic resistance gene nptII using a heat inducible Cre-loxP system from transgenic potato. Plant Mol Biol 62:71–82
Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282
Masumoto C, Miyao M (2009) Agrobacterium-mediated transformation of rice (in Japanese). Teionkagaku 67:641–647
Acknowledgments
This work was supported in part by Promotion of Basic Research Activities for Innovative Biosciences, BRAIN, and JSPS KAKENHI 26292005, Japan.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Murata, M., Kanatani, A., Kashihara, K. (2016). One-Step Generation of Chromosomal Rearrangements in Rice. In: Murata, M. (eds) Chromosome and Genomic Engineering in Plants. Methods in Molecular Biology, vol 1469. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4931-1_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-4931-1_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-4929-8
Online ISBN: 978-1-4939-4931-1
eBook Packages: Springer Protocols