The effect of antibiotics on the shoot regeneration in apple cultivar “Golden Delicious”
2015
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Authors:
Stanišić, MarianaNinković, Slavica
Savić, Jelena
Ćosić, Tatjana
Banjac, Nevena
Contributors
Uzelac, BrankaDocument Type:
Conference object (Published version)
,
© 2015 by the Serbian Plant Physiology Society
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Show full item recordAbstract:
The effect of antibiotics hygromycin, cefotaxime and meropenem on shoot regeneration in apple cultivar “Golden Delicious” was evaluated to optimize the protocol for Agrobacterium tumefaciens-mediated genetic transformation. The careful determination of antibiotics concentration is important step to ensure efficient elimination of Agrobacterium and appropriate selection process without escapes and detrimental effect on plant regeneration. Healthy and fully expanded young leaves of in vitro grown apple shoots were cut
transversely into three pieces that were used as explants for shoot regeneration. Regeneration frequency, the
mean number of shoots per explant and the index of shoot-forming capacity were calculated after 8 weeks
of culturing on Murashige and Skoog medium containing 5 mg L-1 thidiazuron (TDZ) and 0.3 mg L-1 indole-
3-butyric acid (IBA), supplemented with hygromycin (0, 2, 4, 5, 6, 8 and 10 mg L-1), cefotaxime (0, 100, 200,
300, 400, 500 and 600 mg L-1) or meropenem (0, 25, 50, 75, 100, 125 and 150 mg L-1). Results indicated that
the highest index of shoot-forming capacity was obtained at 300 mg L-1 cefotaxime (3.67) comparing to control (2.93) and other antibiotic treatments. The increase of hygromycin concentration significantly decreased
regeneration efficiency, while at concentrations of 5 mg L-1 and higher it completely inhibited shoot regeneration. Therefore, the present study outlines the use of cefotaxime at 300 mg L-1 and a stepwise selection procedure with the final hygromycin concentration of 5 mg L-1 as a suitable system for A. tumefaciens-mediated
genetic transformation of apple cultivar “Golden Delicious”.
Keywords:
cefotaxime; hygromycin; meropenem; transformationFunding / projects:
- Biotechnology in vitro - crop, medicinal and endangered plant species (RS-MESTD-Basic Research (BR or ON)-173015)
In:
- Uzelac B, editor. Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia. Belgrade: Serbian Plant Physiology Society: Institute for Biological Research “Siniša Stanković”; 2015. p. 28-9.