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Light-Dependent Betanin Production by Transformed Cells of Sugar Beet

Dubravko Pavoković1*, Gordana Rusak2, Višnja Besendorfer1 and Marijana Krsnik-Rasol1


1
Department of Molecular Biology, Division of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000 Zagreb, Croatia

2Department of Botany, Division of Biology, Faculty of Science, University of Zagreb, Marulićev trg 9a, HR-10000 Zagreb, Croatia

Article history:

Received July 4, 2008
Accepted December 19, 2008

Key words:

Agrobacterium tumefaciens, betacyanins, carbohydrates, in vitro tissue culture, pigment, secondary metabolites

Summary:

The objective of this work is to transform sugar beet (Beta vulgaris L. var. altissima) cells using Agrobacterium tumefaciens, a wild octopine strain B6S3, and to study metabolic changes associated with the transformation. From the primary tumours on leaf fragments, two tumour lines were established: one pale green and the other red-violet in colour. The red-violet pigment was identified by means of high-performance liquid chromatography as betanin. Betanin production is strongly light-dependent. To enhance the betanin yield, nutrient media with different carbohydrates like sucrose, a combination of glucose and fructose, or only glucose or only fructose were tested. The selection of carbohydrate affected betanin production and yield. It was observed that the production of betanin per g of dry mass was 20–40 % higher on simple carbohydrates, especially fructose, than on sucrose or the combination of glucose and fructose. However, due to higher biomass production on the medium with sucrose, the highest total yield of betanin was obtained in the presence of sucrose. The most suitable time for possible betanin extraction was between days 7 and 14 for tissue grown on sucrose. The tumour line seems to be promising as an alternative source of betanin as well as a model to study sugar-regulated genes involved in tissue morphology control.

 


*Corresponding author:           dubravko@zg.biol.pmf.hr
                                               ++385 (0)1 4606 458
                                               ++385 (0)1 4606 286

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