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An improved rice transformation system using the biolistic method

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Abstract

Immature embryos and embryogenic calli of rice, both japonica and indica subspecies, were bombarded with tungsten particles coated with plasmid DNA that contained a gene encoding hygromycin phosphotransferase (HPH, conferring hygromycin resistance) driven by the CaMV 35S promoter or Agrobactenum tumefaciens NOS promoter. Putatively transformed cell clusters were identified from the bombarded tissues 2 weeks after selection on hygromycin B. By separating these cell clusters from each other, and by stringent selection not only at the callus growth stage but also during regeneration and plantlet growth, the overall transformation and selection efficiencies were substantially improved over those previously reported. From the most responsive cultivar used in these studies, an average of one transgenic plant was produced from 1.3 immature embryos or from 5 pieces of embryogenic calli bombarded. Integration of the introduced gene into the plant genome, and inheritance to the offspring were demonstrated. By using this procedure, we have produced several hundred transgenic plants. The procedure described here provides a simple method for improving transformation and selection efficiencies in rice and may be applicable to other monocots.

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Abbreviations

bp:

base pairs

CaMV:

cauliflower mosaic virus

GUS:

β-glucuronidase

HPH:

hygromycin phosphotransferase

hyg B:

hygromycin B

hygr :

hygromycin resistance

NOS:

Agrobactenum tumefaciens nopaline synthase

PCR:

polymerase chain reaction

X-Gluc:

5-bromo-4-chloro-3-indolyl-β-D-glucuronide

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Authors and Affiliations

  1. Institute of Genetics, Academia Sinica, Beijing, China

    Liangcai Li

  2. International Laboratory for Tropical Agricultural Biotechnology (ILTAB), Division of Plant Biology, Department of Cell Biology, The Scripps Research Institute, 92037, La Jolla, CA, USA

    Rongda Qu, Alexandre de Kochko, Claude Fauquet & Roger N. Beachy

Authors
  1. Liangcai Li
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  2. Rongda Qu
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  3. Alexandre de Kochko
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  4. Claude Fauquet
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  5. Roger N. Beachy
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Communicated by G. C. Phillips

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Li, L., Qu, R., de Kochko, A. et al. An improved rice transformation system using the biolistic method. Plant Cell Reports 12, 250–255 (1993). https://doi.org/10.1007/BF00237129

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  • DOI: https://doi.org/10.1007/BF00237129

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