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A modified method for routine Agrobacterium-mediated transformation of in vitro grown potato microtubers

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Summary

In vitro-grown potato (Solanum tuberosum L.) microtubers were used as an explant source in the production of transgenic plants by Agrobacterium-mediated gene transfer. In this study we tested four diverse potato cultivars, Lemhi Russet, Russet Burbank, Wauseon, and Yankee Chipper on various levels of zeatin riboside and 3-indoleacetyl-DL-aspartic acid for their ability to regenerate transgenic plants after infection with Agrobacterium tumefaciens. Culturing microtuber blocks from the medullary area separately from cortex and epidermal tissue containing the eyes resulted in fewer transgenic plants, with transgenic shoots arising only from the tissue with the eyes. Lemhi and Russet Burbank microtuber discs were also transformed with a chimeric gene, CLaSP, designed to increase resistance to blackspot bruise in the tuber. This method resulted in transformed plants in every experiment, with an efficiency that appeared to be genotype dependent.

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Abbreviations

GUS:

β-glucuronida (uidA)

IAA-AA:

3-indoleacetyl-DL-aspartic acid

LB:

Luria-Bertani

LSP:

larval serum storage protein

nos:

nopaline synthase

npt II:

neomycin phosphotransferase

MS:

Murashige and Skoog

PHA:

phytohemaglutinin

ZR:

zeatin riboside

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Authors and Affiliations

  1. Western Regional Research Center, U.S.Department of Agriculture, Agricultural Research Service, 800 Buchanan Street, 94710, Albany, CA, USA

    Gordon W. Snyder & William R. Belknap

Authors
  1. Gordon W. Snyder
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  2. William R. Belknap
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Communicated by J. M. Widholm

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Snyder, G.W., Belknap, W.R. A modified method for routine Agrobacterium-mediated transformation of in vitro grown potato microtubers. Plant Cell Reports 12, 324–327 (1993). https://doi.org/10.1007/BF00237428

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  • DOI: https://doi.org/10.1007/BF00237428

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