Abstract
The high-capacity binary bacterial artificial chromosome (BIBAC) vector system permits the insertion of large fragments of DNA, up to 150 kb, into plants via Agrobacterium-mediated transformation. Here, we describe an optimized protocol for transformation of japonica rice (Oryza sativa L.) using this system. Calli derived from mature embryos are transformed using Agrobacterium strain LBA4404 that carries the BIBAC vector and the super-virulent helper plasmid pCH32. Transformed calli are then regenerated using optimized media and tested for transgene integration by PCR, GUS assay, and Southern blot analyses.
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
Similar content being viewed by others
References
Hellens R, Mullineaux P, Klee H (2000) A guide to Agrobacterium binary Ti vectors. Trends Plant Sci 5:446–451
Ercolano MR, Ballvora A, Paal J, Steinbiss HH, Salamini F, Gebhardt C (2004) Functional complementation analysis in potato via biolistic transformation with BAC large DNA fragments. Mol Breeding 13:15–22
Hamilton CM (1997) A binary-BAC system for plant transformation with high-molecular-weight DNA. Gene 200:107–116
Hamilton CM, Frary A, Lewis C, Tanksley SD (1996) Stable transfer of intact high molecular weight DNA into plant chromosome. Proc Natl Acad Sci U S A 93:9975–9979
Frary A, Hamilton CM (2001) Efficiency and stability of high molecular weight DNA transformation: an analysis in tomato. Transgenic Res 10:121–132
Hamilton CM, Frary A, Xu Y, Tanksley SD, Zhang HB (1999) Construction of tomato genomic DNA libraries in a binary-BAC (BIBAC) vector. Plant J 18:223–229
He RF, Wang Y, Shi Z, Ren X, Zhu L, Weng Q, He GC (2003) Construction of a genomic library of wild rice and Agrobacterium-mediated transformation of large insert DNA linked to BPH resistance locus. Gene 321:113–121
Wang W, Wu Y, Li Y, Xie J, Zhang Z, Deng Z, Zhang Y, Yang C, Lai J, Zhang H, Bao H, Tang S, Yang C, Gao P, Xia G, Guo H, Xie Q (2010) A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes. Plant Mol Biol 72:91–99
Shibata D, Liu YG (2000) Agrobacterium-mediated plant transformation with large DNA fragments. Trends Plant Sci 5:354–357
Naqvi S, Farre G, Sanahuja G, Capell T, Zhu C, Christou P (2010) When more is better: multigene engineering in plants. Trends Plant Sci 15:48–56
Osbourn A (2010) Secondary metabolic gene clusters: evolutionary toolkits for chemical innovation. Trends Genet 26:449–457
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
He RF, Wang YY, Du B, Tang M, You AQ, Zhu LL, He GC (2006) Development of transformation system of rice based on binary bacterial artificial chromosome (BIBAC) vector. Acta Genet Sin 33:269–276
McCouch SR, Kochert G, Yu ZH, Wang ZY, Khush GS, Coffman WR, Tanksley SD (1988) Molecular mapping of rice chromosome. Theor Appl Genet 76:815–829
He R, Pan J, Zhu L, He G (2010) Agrobacterium-mediated transformation of large DNA fragments using a BIBAC vector system in rice. Plant Mol Biol Rep 28:613–619
Acknowledgements
We thank the Plant Science Center of Cornell University for providing the BIBAC2 vector and pCH32 plasmid. This work was supported by the National Natural Science Foundation of China (No. 30470922), the Natural Science Foundation of Hubei Province (No. 2004ABA117), and the National Program of High Technology Development of China (No. 2004AA227120).
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
He, R. (2016). Multigene Engineering in Rice Using High-Capacity Agrobacterium tumefaciens BIBAC Vectors. In: MacDonald, J., Kolotilin, I., Menassa, R. (eds) Recombinant Proteins from Plants. Methods in Molecular Biology, vol 1385. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3289-4_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3289-4_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3288-7
Online ISBN: 978-1-4939-3289-4
eBook Packages: Springer Protocols