Abstract
The effect, on adventitious regeneration from apricot leaf explants and transformation of leaf tissues, of auxins pulses with NAA and 2, 4-D was tested. Addition of the polyamines putrescine and spermidine to the regeneration medium, alone or in combination with the ethylene inhibitors silver thiosulphate and aminoethoxyvinylglycine, were also tested to design a procedure that improved transformation efficiency. Spermidine at 2 mM in combination with 0.5 μM aminoethoxyvinylglycine and four-day pulses with two different concentrations of 2, 4-D increased significantly shoot regeneration. Spermidine at the same concentration but in combination with 60 μM silver thiosulphate and four-day pulses with 9 μM 2, 4-D also increased stable transformation events and GFP-expressing calluses probably by inducing a larger amount of dividing cells where Agrobacterium transferred its T-DNA. Since regeneration from apricot leaves occurs mostly from developing calluses, it is important to obtain many GFP-expressing calluses and, given that transformation efficiencies (number of transformed shoots per total number of explants) in woody plants are generally very low, approaches that allow the optimization of T-DNA transfer and total number of transformed cells obtained, will improve probabilities of obtaining transformed shoots.
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Abbreviations
- AS:
-
acetosyringone
- AVG:
-
aminoethoxyvinylglycine
- DKW:
-
Driver and Kuniyuki (1984)
- GFP:
-
green fluorescent protein
- QL:
-
Quoirin and Lepoivre (1977)
- NAA:
-
naphthaleneacetic acid
- Put:
-
putrescine
- Spd:
-
spermidine
- Spm:
-
spermine
- STS:
-
silver thiosulphate
- TDZ:
-
thidiazuron
- 2, 4-D:
-
2, 4 dichlorophenoxy –acetic acid
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Petri, C., Alburquerque, N., Pérez-Tornero, O. et al. Auxin pulses and a synergistic interaction between polyamines and ethylene inhibitors improve adventitious regeneration from apricot leaves and Agrobacterium-mediated transformation of leaf tissues. Plant Cell Tiss Organ Cult 82, 105–111 (2005). https://doi.org/10.1007/s11240-004-7013-y
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DOI: https://doi.org/10.1007/s11240-004-7013-y