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Agrobacterium-mediated transformation of quaking aspen (Populus tremuloides) and regeneration of transgenic plants

Plant Cell Reports volume 14, pages 94–97 (1994)Cite this article

Summary

Agrobacterium-mediated gene transformation of Populus tremuloides Michx was accomplished by co-cultivation of leaf disks excised from greenhouse plants with Agrobacterium tumefaciens containing a binary Ti-plasmid vector harboring chimeric neomycin phosphotransferase (NPT II) and ß-glucuronidase (GUS) genes. Shoot regeneration in the presence of kanamycin was achieved when thidiazuron (TDZ) was used as a plant growth regulator. Transformation was verified by amplification of NPT II and GUS gene fragments from genomic DNA of transgenic plants with polymerase chain reaction (PCR) and integration of these genes into nuclear genome of transgenic plants was confirmed by genomic Southern hybridization analysis. Histochemical assay revealed the expression of GUS gene in leaf, stem and root tissues of transgenic plants, further confirming the integration and expression of T-DNA in these plants. This protocol allows effective transformation and regeneration of quaking aspen using greenhouse-grown materials as an explant source. Whole plant regeneration from cuttings of fieldgrown mature quaking aspen and hybrid poplar (P. alba x P. grandidentata) was also readily achieved by using this protocol, which represents a potential system for producing transgenic quaking aspen and hybrid poplar of valuable genotypes.

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Abbreviations

AMV RNA4:

Alfalfa mosaic virus RNA4

BA:

6-benzyladenine

CaMV:

cauliflower mosaic virus

2,4-D:

2,4-dichlorophenoxyacetic acid

EDTA:

ethylenediaminetetraacetic acid

FAA:

formalin-acetic acid-alcohol

GUS:

ß-glucuronidase

NAA:

1-naphthylacetic acid

NPT II:

neomycin phosphotransferase II

PCR:

polymerase chain reaction

SDS:

sodium dodecyl sulphate

TE:

Tris-Cl/EDTA

TDZ:

N-phenyl-N′-1,2,3-thiadiazol-5-yl-urea (thidiazuron)

WPM:

woody plant medium (Lloyd and McCown 1980)

X-GLUC:

5-bromo-4-chloro-3-indolyl-ß-glucuronic acid

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Author information

Affiliations

  1. School of Forestry and Wood Products, Michigan Technological University, 49931, Houghton, Michigan, USA

    Chung-Jui Tsai & Vincent L. Chiang

  2. Department of Biological Sciences, Michigan Technological University, 49931, Houghton, Michigan, USA

    Gopi K. Podila

Authors
  1. Chung-Jui Tsai
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  2. Gopi K. Podila
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  3. Vincent L. Chiang
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Additional information

Communicated by J. Widholm

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Tsai, CJ., Podila, G.K. & Chiang, V.L. Agrobacterium-mediated transformation of quaking aspen (Populus tremuloides) and regeneration of transgenic plants. Plant Cell Reports 14, 94–97 (1994). https://doi.org/10.1007/BF00233768

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  • DOI: https://doi.org/10.1007/BF00233768

Keywords

  • Transgenic Plant
  • Neomycin
  • Thidiazuron
  • Hybrid Poplar
  • Neomycin Phosphotransferase