A protocol for sonication-assisted Agrobacterium rhizogenes-mediated transformation of haploid and diploid sugar beet (Beta vulgaris L.) explants

• Authors: Magdalena Klimek-Chodacka , Rafal Baranski
• Languages of publication: EN

Abstracts

EN

Hairy root cultures obtained after Agrobacterium rhizogenes-mediated genetic transformation can serve as a model system for studying plant metabolism and physiology, or can be utilized for the production of secondary metabolites. So far no efficient protocol of hairy root development in sugar beet has been publically released. In this work, two A. rhizogenes strains (A4T and LBA1334) carrying a binary vector pBIN-m-gfp5-ER or pCAMBIA1301 possessing gfp and uidA reporter genes were used to transform petiole explants of haploid and diploid sugar beet genotypes. Five treatment combinations of sonicated-assisted Agrobacterium-mediated transformation were compared. Hairy roots appeared on 0% to 54% of explants depending on the treatment combination used. The highest frequency was achieved when explants of a diploid genotype were sonicated for 15 s in the inoculum containing A. rhizogenes of OD600=0.5 and then co-cultured for three days. Using the same treatment combinations the explants of haploid genotypes developed hairy roots with the frequency ranging from 10% to 36%. Transformation efficiency was independent on the bacterial strain used. The results indicate that haploid sugar beet explants are amenable to transformation using A. rhizogenes, and that the efficiency of that process can be increased by applying short ultrasound treatment.

Keywords

EN

genetic modification; hairy root; SAAT; ultrasound

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2014
• Volume: 61
• Issue: 1
• Pages: 13-17

Dates

published

2014

received

2013-10-15

revised

2014-03-11

accepted

2014-03-17

(unknown)

2014-03-22

Contributors

author

Magdalena Klimek-Chodacka

• Institute of Plant Biology and Biotechnology, Faculty of Horticulture, University of Agriculture in Krakow, Kraków, Poland

author

Rafal Baranski

• Institute of Plant Biology and Biotechnology, Faculty of Horticulture, University of Agriculture in Krakow, Kraków, Poland

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