Advertisement

Rice, Indica (Oryza sativa L.)

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1223)

Abstract

Indica varieties, which are generally recalcitrant to tissue culture and transformation, occupy 80 % of rice cultivation area in the world. Therefore, transformation method for indica rice must be improved greatly so that global rice production would take full advantage of cutting-edge biotechnology. An efficient protocol for indica transformation mediated by Agrobacterium tumefaciens is hereby described. Immature embryos collected from plants in a greenhouse are cocultivated with A. tumefaciens after pretreatment with heat and centrifuging. The protocol was successfully tested in many elite indica cultivars such as IR8, IR24, IR58025B, IR64, IR72, Suweon 258, and Nanjing 11, yielding between 5 and 15 of independent transgenic plants per immature embryo. The use of immature embryos is recommended because gene transfer to them could be much more efficient and much less genotype dependent than gene transfer to callus.

Key words

Agrobacterium tumefaciens Indica Rice Transformation 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgment

We thank Ms. Chiaki Sakuma and Ms. Tomoko Sakai for technical assistance.

References

  1. 1.
    Glaszmann JC (1987) Isozymes and classification of Asian rice varieties. Theor Appl Genet 74:21–30CrossRefPubMedGoogle Scholar
  2. 2.
    Garris AJ, Tai TH, Coburn J, Kresovich S, McCouch S (2005) Genetic Structure and Diversity in Oryza sativa L. Genetics 169:1631–1638CrossRefPubMedCentralPubMedGoogle Scholar
  3. 3.
    Hiei Y, Ishida Y, Kasaoka K, Komari T (2006) Improved frequency of transformation in rice and maize by treatment of immature embryos with centrifugation and heat prior to infection with Agrobacterium tumefaciens. Plant Cell Tissue Organ Cult 87:233–243CrossRefGoogle Scholar
  4. 4.
    Hiei Y, Komari T (2006) Improved protocols for transformation of indica rice mediated by Agrobacterium tumefaciens. Plant Cell Tiss Organ Cult 85:271–283CrossRefGoogle Scholar
  5. 5.
    Kim DS, Lee IS, Jang CS, Hyun DY, Seo YW, Lee YI (2004) Selection of 5-methyltryptophan resistant rice mutants from irradiated calli derived from embryos. Euphytica 135:9–19CrossRefGoogle Scholar
  6. 6.
    Park MC, Shin J, Kim N, Cho H, Park S, An K, Lee S, An G (2003) High-frequency Agrobacterium-mediated genetic transformation of Tongil rice varieties. J Plant Biol 46:23–30CrossRefGoogle Scholar
  7. 7.
    Rachmawati D, Hosaka T, Inoue E, Anzai H (2004) Agrobacterium-mediated transformation of Javanica rice cv. Rojolele. Biosci Biotechnol Biochem 68:1193–1200CrossRefPubMedGoogle Scholar
  8. 8.
    Hoque ME, Mansfield JW, Bennett MH (2005) Agrobacterium-mediated transformation of indica rice genotypes: an assessment of factors affecting the transformation efficiency. Plant Cell Tiss Organ Cult 82:45–55CrossRefGoogle Scholar
  9. 9.
    Kumar KK, Maruthasalam S, Loganathan M, Sudhakar D, Balasubramanian P (2005) An improved Agrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars. Plant Mol Biol Rep 23:67–73CrossRefGoogle Scholar
  10. 10.
    Lin YJ, Zhang Q (2005) Optimising the tissue culture conditions for high efficiency transformation of indica rice. Plant Cell Rep 23:540–547CrossRefPubMedGoogle Scholar
  11. 11.
    Sahoo K, Tripathi A, Pareek A, Sopory S, Singla-Pareek S (2011) An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars. Plant Methods 7:1–11CrossRefGoogle Scholar
  12. 12.
    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 6:271–282CrossRefPubMedGoogle Scholar
  13. 13.
    Ohta S, Mita S, Hattori T, Nakamura K (1990) Construction and expression in tobacco of a β-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31:805–813Google Scholar
  14. 14.
    Toriyama K, Arimoto Y, Uchimiya H, Hinata K (1988) Transgenic plants after direct gene transfer into protoplasts. Bio/Technology 6:1072–1074CrossRefGoogle Scholar
  15. 15.
    Lucca P, Ye X, Potrykus I (2001) Effective selection and regeneration of transgenic rice plants with mannose as selective agent. Mol Breed 7:43–49CrossRefGoogle Scholar
  16. 16.
    Huang JQ, Wel ZM, An HL, Zhu YX (2001) Agrobacterium tumefaciens-mediated transformation of rice with the spider insecticidal gene conferring resistance to leaf folder and striped stem borer. Cell Res 11:149–155CrossRefPubMedGoogle Scholar
  17. 17.
    Christou P, Ford TL, Kofron M (1991) Production of transgenic rice (Oryza sativa L.) from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/Technology 9:957–962CrossRefGoogle Scholar
  18. 18.
    Enriquez-Obregon GA, Prieto-Samsonov DL, de la Riva GA, Perez M, Selman-Housein G, Vazquez-Padron RI (1999) Agrobacterium-mediated Japonica rice transformation: a procedure assisted by an antinecrotic treatment. Plant Cell Tiss Organ Cult 59:159–168CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Plant Innovation CenterJapan Tobacco Inc.IwataJapan

Personalised recommendations

Cite protocol

Buy options