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Transgenic poplars: expression of chimeric genes using four different constructs

Summary

Leaf or stem explants of a hybrid poplar clone (Populus tremula X Populus alba), sensitive to Agrobacterium tumefaciens, were co-cultivated either by an octopine or a nopaline disarmed A. tumefaciens modified strain. Transformed poplar shoots were readily regenerated from explants. The protocol was improved using the nopaline disarmed strain C58/pMP90 with the binary vector pBI121. This protocol was then used to test three other vectors. The first one, possessing a nptII gene fused to the CaMV 19S promoter, permitted regeneration of transformed shoots in presence of 50 to 100 mg/l kanamycin. The two other vectors carried an additional nptII gene under the control of the CaMV 35S or CaMV 35S promoter with a double enhancer sequence (CaMV 70). CaMV 70 promoter provided consistently higher level of gene expression than the other promoters in both callus and leaf tissues.

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Abbreviations

CaMV:

Cauliflower Mosaïc Virus

2iP:

2-isopentenyladenine

GUS and gus :

ß-glucuronidase

NAA:

1-naphthaleneacetic acid

NPTII and nptII :

neomycin phosphotransferase II

NOS:

Nopaline synthase, X-Gluc: 5-bromo-4-chloro-3-indolyl ß-D glucuronide

Ap:

ampicillin

Gn:

gentamycin

Km:

kanamycin

Rf:

rifampicin

St:

streptomycin

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Additional information

This work is dedicated to the late Marie France Michel who initiated the poplar biotechnology project at INRA.

Communicated by N. Amrhein

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Leple, J.C., Brasileiro, A.C.M., Michel, M.F. et al. Transgenic poplars: expression of chimeric genes using four different constructs. Plant Cell Reports 11, 137–141 (1992). https://doi.org/10.1007/BF00232166

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Keywords

  • Alba
  • Agrobacterium Tumefaciens
  • Hybrid Poplar
  • Chimeric Gene
  • nptII Gene