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Piercing and vacuum infiltration of the mature embryo: a simplified method for Agrobacterium-mediated transformation of indica rice

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

Traditional transformation methods are complex and time consuming. It is generally difficult to transform indica rice varieties using traditional transformation methods due to their poor regeneration. In this contribution, a simple method was developed for the transformation of indica rice. In this method, the mature embryos of soaked seeds were pierced by a needle, and then soaked in the Agrobacterium inoculum under vacuum infiltration. The inoculated seeds germinated and grew to maturation (T 0) under nonsterile conditions. The herbicide or antibiotic analysis and molecular analysis were conducted on T 0 plants. The results showed that although the efficiency of transformation was about 6.0%, it was easier to transform indica rice using the proposed method, and the transformation process was significantly shortened. The success of transformation was further confirmed by the genetic and molecular analyses of T 1 transformants.

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Abbreviations

6-BA:

6-Benzylaminopurine

CTAB:

Cetyltrimethylammonium bromide

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

MS:

Murashige and Skoog

PCR:

Polymerase chain reaction

YEB:

Yeast extract–beef extract medium

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Acknowledgments

The authors acknowledge the financial support of the Key Science and Technology Financing Projects of Ministry of Education of China (No: 2002-78).

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Authors and Affiliations

  1. Institute of Bioscience and Biotechnology, Hunan University, Changsha, 410082, Hunan, China

    Jianzhong Lin, Bo Zhou, Jin Mei, Xiaoying Zhao, Xinhong Guo, Xingqun Huang, Dongying Tang & Xuanming Liu

  2. Academy of Seed Industry of Hunan Yahua, Changsha, 410001, Hunan, China

    Yuanzhu Yang

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  1. Jianzhong Lin
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  2. Bo Zhou
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  3. Yuanzhu Yang
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  4. Jin Mei
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  5. Xiaoying Zhao
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  6. Xinhong Guo
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  7. Xingqun Huang
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  8. Dongying Tang
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  9. Xuanming Liu
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Corresponding author

Correspondence to Xuanming Liu.

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Communicated by W. Harwood.

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Lin, J., Zhou, B., Yang, Y. et al. Piercing and vacuum infiltration of the mature embryo: a simplified method for Agrobacterium-mediated transformation of indica rice. Plant Cell Rep 28, 1065–1074 (2009). https://doi.org/10.1007/s00299-009-0706-2

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  • DOI: https://doi.org/10.1007/s00299-009-0706-2

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