Abstract
Genetic transformation plays a key role in deciphering regulation of agronomic traits at molecular level in rice, a model monocot cereal crop. Here we describe an efficient and fast protocol for producing transgenic japonica rice plants using the Agrobacterium-mediated transformation method. The protocol simplifies medium compositions and transformation steps and can be easily followed by a lab technician with little tissue culture experience. Using this protocol, we have transformed thousands of gene constructs in the past 10 years and edited hundreds of genes with the CRISPR-Cas9 system recently.
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References
Shimamoto K, Terada R, Izawa T, Fujimoto H (1989) Fertile transgenic rice plants regenerated from transformed protoplasts. Nature 338:274–276
Christou P, Ford T, Kofron M (1991) Production of transgenic rice (Oryza Sativa L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Nat Biotechnol 9:957–962
Dai S, Zheng P, Marmey P, Zhang S, Tian W, Chen S, Beachy RN, Fauquet C (2001) Comparative analysis of transgenic rice plants obtained by Agrobacterium-mediated transformation and particle bombardment. Mol Breed 7:25–33
Chen L, Marmey P, Taylor NJ, Brizard JP, Espinoza C, D'Cruz P, Huet H, Zhang S, Kochko A, Beachy RN, Fauquet CM (1998) Expression and inheritance of multiple transgenes in rice plants. Nat Biotechnol 16:1060–1064
Zhu C, Naqvi S, Breitenbach G, Sandmann J, Christou P, Capell T (2008) Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize. Proc Natl Acad Sci U S A 105:18232–18237
Naqvi S, Zhu C, Farre G, Bassie L, Ramessar K, Breitenbach J, Perez-Conesa D, Ros-Berruezo G, Sandmann G, Capell T, Christou P (2009) Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways. Proc Natl Acad Sci U S A 106:7762–7767
Zhang Y, Liang Z, Zong Y, Wang Y, Liu J, Chen K, Qiu JL, Gao C (2016) Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA. Nat Commun 7:12617. https://doi.org/10.1038/ncomms12617
Svitashev S, Schwartz C, Lenderts B, Young JK, Cigan AK (2016) Genome editing in maize directed by CRISPR–Cas9 ribonucleoprotein complexes. Nat Commun 7:13274. https://doi.org/10.1038/ncomms13274
Liang Z, Chen K, Li T, Zhang Y, Wang Y, Zhao Q, Liu J, Zhang H, Liu C, Ran Y, Gao C (2017) Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nat Commun 8:14261. https://doi.org/10.1038/ncomms14261
Rained DM, Bottino P, Gordon MP, Nester EW (1990) Agrobecterium-mediated transformation of rice (Oryza sativa L.). Nat Biotechnol 8:33–38
Chan MT, Chang HH, Ho SL, Tong WF, Yu SM (1993) Agrobacteriummediated production of transgenic rice plants expressing a chimeric ɑ-amylase promoter/β glucuronidase gene. Plant Mol Biol 22:491–506
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
Hiei Y, Komari T (2008) Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nat Protoc 3:824–834
Roy M, Jain RK, Rohila JS, Wu R (2000) Production of agronomically superior transgenic rice plants using Agrobacterium transformation methods: present status and future perspectives. Curr Sci 79:954–960
Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H (2006) Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. Plant J 47:969–976
Zhou F, Lin Q, Zhu L et al (2013) D14–SCFD3-dependent degradation of D53 regulates strigolactone signaling. Nature 504:406–410
Gao H, Jin M, Zheng X et al (2014) Days to heading 7, a major quantitative locus determining photoperiod sensitivity and regional adaptation in rice. Proc Natl Acad Sci U S A 46:16337–16342
Liu Y, Wu H, Chen H et al (2015) A gene cluster encoding lectin receptor kinases confers broad-spectrum and durable insect resistance in rice. Nat Biotechnol 33:301–305
Wu S, Xie Y, Zhang J et al (2015) VLN2 regulates plant architecture by affecting microfilament dynamics and polar auxin transport in rice. Plant Cell 27:2829–2845
Sun Y, Zhang X, Wu C, He Y, Ma Y, Hou H, Guo X, Du W, Zhao Y, Xia L (2016) Engineering herbicide-resistant rice plants through CRISPR/Cas9-mediated homologous recombination of acetolactate synthase. Mol Plant 9:628–631
Acknowledgment
This work was supported by the Innovation Program of Chinese Academy of Agricultural Sciences.
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Wu, C., Sui, Y. (2019). Efficient and Fast Production of Transgenic Rice Plants by Agrobacterium-Mediated Transformation. In: Kumar, S., Barone, P., Smith, M. (eds) Transgenic Plants. Methods in Molecular Biology, vol 1864. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8778-8_7
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DOI: https://doi.org/10.1007/978-1-4939-8778-8_7
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