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The effect of aminoglycoside antibiotics on the adventitious regeneration from apricot leaves and selection of nptII-transformed leaf tissues

Abstract

The effect of different concentrations of the aminoglycoside antibiotics, geneticin, paromomycin and streptomycin on adventitious regeneration from leaf explants of apricot was tested to design an alternative procedure for selecting transgenic shoots. Streptomycin and paromomycin reduced shoot regeneration percentage with increasing concentration of antibiotics. Almost a complete inhibition of regeneration was reached when 20μM paromomycin was used, although up to 40 μM streptomycin was necessary to completely inhibit regeneration. Geneticin had a very toxic effect on apricot leaves and regeneration was inhibited at almost all concentrations tested. Addition of kanamycin hastened the development of adventitious buds although silver thiosulfate and not kanamycin was responsible for the observed increase in the consistency of the results from independent experiments. Kanamycin and paromomycin at the concentrations tested improved selection of transformed cells and resulted in a larger number of gfp-expressing regions. Paromomycin at 40 and 25.7 μM kanamycin improved proliferation of transformed tissues as compared with the other antibiotics used and non-selected controls.

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Abbreviations

AS:

acetosyringone

CV:

coefficient of variation

DKW:

Driver and Kuniyuki (1984)

GFP:

green fluorescent protein

NAA:

naphthaleneacetic acid

QL:

Quoirin and Lepoivre (1977)

STS:

silver thiosulphate

TDZ:

thidiazuron

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Correspondence to Lorenzo Burgos.

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Petri, C., Alburquerque, N. & Burgos, L. The effect of aminoglycoside antibiotics on the adventitious regeneration from apricot leaves and selection of nptII-transformed leaf tissues. Plant Cell Tiss Organ Cult 80, 271–276 (2005). https://doi.org/10.1007/s11240-004-1019-3

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Keywords

  • adventitious regeneration
  • Agrobacterium-mediated transformation
  • Prunus armeniaca L
  • selective marker