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Regeneration of fertile transgenic indica (group 1) rice plants following microprojectile transformation of embryogenic suspension culture cells

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Abstract

Regenerable embryogenic suspensions of elite Indica (group 1) rice varieties IR24, IR64, IR72 and an advanced Indica rice breeding line IR57311-95-2-3 were established within 6–8 weeks from 3–4 week old calli derived from mature seeds. Transgenic rice plants were obtained by introducing a plasmid carrying genes encoding hygromycin phosphotransferase (hph, conferring resistance to hygromycin B) and ß-glucuronidase (uidA), both driven by the CaMV 35S promoter, via particle bombardment of embryogenic suspensions. The effect of osmotic conditioning on transformation was evaluated. Regenerated plants were resistant to hygromycin B and expressed the uidA (GUS) gene. The growth of mother plants (R0) was normal and seeds were produced. Southern blot analysis of R0 and R1 plants showed that hygromycin resistant plants contained intact hph genes that were inherited in a Mendelian fashion. A protocol for a simple, efficient, repeatable, genotype- and environment-independent Indica rice transformation system is described.

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Abbreviations

2,4-D:

2,4-dichlorophenoxy acetic acid

NAA:

α-naphthalene acetic acid

kb:

kilobase

GUS:

ß-glucuronidase

hph:

hygromycin B phosphotransferase

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Authors and Affiliations

  1. Division of Plant Biology, ILTAB/TSRI-ORSTOM — The Scripps Research Institute, 10666 North Torrey Pines Road — MRC7, 92037, La Jolla, CA, USA

    Shiping Zhang, Lili Chen, Rongda Qu, Philippe Marmey, Roger Beachy & Claude Fauquet

Authors
  1. Shiping Zhang
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  2. Lili Chen
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  3. Rongda Qu
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  4. Philippe Marmey
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  5. Roger Beachy
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  6. Claude Fauquet
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Communicated by I. K. Vasil

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Zhang, S., Chen, L., Qu, R. et al. Regeneration of fertile transgenic indica (group 1) rice plants following microprojectile transformation of embryogenic suspension culture cells. Plant Cell Reports 15, 465–469 (1996). https://doi.org/10.1007/BF00232975

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  • DOI: https://doi.org/10.1007/BF00232975

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