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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Molecular Plant Pathology Laboratory » Research » Publications at this Location » Publication #311223

Title: Gentiana dinarica Beck hairy root cultures and evaluation of factors affecting growth and xanthone production

Author
item VINTERHALTER, BRANKA - University Of Belgrade
item DIJANA, KRSTIC-MILOSEVI - University Of Belgrade
item TEODORA, JANKOVIC - Institut Za Proucavanje Lekovitog Bilja
item DEJAN, PLJEVLJAKUSIC - Institut Za Proucavanje Lekovitog Bilja
item NINKOVIC, SLAVICA - University Of Belgrade
item Smigocki, Anna
item VINTERHALTER, DRAGAN - University Of Belgrade

Submitted to: Plant Cell Tissue and Organ Culture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/10/2015
Publication Date: 2/20/2015
Citation: Vinterhalter, B., Dijana, K.C., Teodora, J., Dejan, P., Ninkovic, S., Smigocki, A.C., Vinterhalter, D. 2015. Gentiana dinarica Beck hairy root cultures and evaluation of factors affecting growth and xanthone production. Plant Cell Tissue And Organ Culture. DOI: 10.1007/s11240-015-0737-z.

Interpretive Summary: Gentians are sub-alpine, perennial plants used in traditional medicines for preparation of bitter tonic mainly used as a remedy for digestive problems. Most recently, the active compounds (xanthones) in Gentians have been recognized as having hepatoprotective, anti-inflammatory and antidiabetic properties. These active compounds are produced in roots and have been found in only two other plants in the world. To better understand how xanthones are made and to speed up the production process of these active compounds, we developed and optimized a method for growing Gentian roots in the laboratory. The laboratory grown roots offer an approach that scientists will use to develop methods for rapid production of large quantities of essentially pure bioactive xanthones.

Technical Abstract: The induction and establishment of hairy root cultures of Gentiana dinarica using two strains of Agrobacterium rhizogenes (A4M70GUS and 15834/PI) is reported for the first time. Hairy roots were formed from the shoots 25 days after inoculation, and strain 15834/PI had higher induction rate of hairy roots than strain A4M70GUS. Hairy roots were cultured in plant growth regulator free ½ MS liquid medium and 14 clones were obtained. Transgenic status of the roots was confirmed by GUS assay and PCR analysis. Root clones were characterized on the basis of growth pattern and type of branching, and two clones per strain were studied for their growth parameters and phenolic production. A two level full factorial design was used to determine the dependence of the factors such as carbohydrate type, carbohydrate concentration, and type of clone, as well as their interactions in affecting the dry weight, total phenolic content and norswertianin-1-O-primeveroside production in hairy root cultures. The results showed that carbohydrate type and concentration had the most significant effect on the dry weight of root, while carbohydrate concentration caused significant effect on the total phenolic content and norswertianin-1-O-primeveroside production. Clones D and 3 cultured on 116.8 mM sucrose concentration could be recommended for the optimal conditions in obtaining high root biomass and phenolic production.