Improved podophyllotoxin production by transformed cultures of Linum album

Abstract

Various cell and hairy root cultures of L. album were developed and analyzed for podophyllotoxin content. Transformed callus and hairy root cultures developed from infection of stem portions of in vitro-germinated L. album plant with Agrobacterium rhizogenes NCIM 5140 strain were selected on the basis of high podophyllotoxin content and growth. Based on the integration of Ri T(L)-DNA and T(R)-DNA, integration of only the ags and not the rol gene in transformed cell culture indicated fragmented integration pattern. The effect of different cultivation media and carbon source on growth and podophyllotoxin production were studied in shake-flask suspension cultures. Detailed batch growth and production kinetics with sugar consumption profile were also established. Maximum volumetric productivity of 4.40 and 2.75 mg/L per day was obtained in cell suspension and hairy root cultures, respectively.

Enhanced production of an anti-malarial compound artesunate by hairy root cultures and phytochemical analysis of Artemisia pallens Wall.

“…Transformation efficiency depends upon type of explants and strain of A. rhizogenes . However, NCIM 5140 strain of A. rhizogenes was widely used for hairy root induction in many plants such as Linum album (Baldi et al 2008), Abrus precatorius L. (Karwasara and Dixit 2009), Helianthus annuus L. (Jha et al 2013) and Sesuvium portulacastrum L. (Lokhande et al 2015). …”

Optimization of podophyllotoxin extraction method fromLinum albumcell cultures

“…We compared the effectiveness of the different reported methods for the extraction of PTOX from plant material (Canel et al, 2001;Van Uden et al, 1989;Zhang et al, 2007;Baldi et al, 2008;Chattopadhyay et al, 2004), applying them to Linum cell cultures. As can be seen in Figure 3, the best results were obtained by using methanol/dichloromethane, followed by the buffer method.…”

“…The supernatant was removed after centrifugation (5000 rpm, 10 min) and evaporated to dryness. The extract was redissolved in HPLC grade methanol and then subjected to analysis by HPLC (Baldi et al, 2008).…”

Lignan formation in hairy root cultures of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter)

“…With regard to the production of lignans from hairy roots a number of studies have focused on podophyllotoxin and its derivatives, an important lead for anticancer drugs. For example, hairy root lines of Linum album produced 105 μg/L [19] or 5.12 mg/L [20] of the lignan while a content of 14.11 mg/L was measured in the roots of wild growing plants [19]. In Linum flavum , hairy roots were reported to contain up to 3.5% 5-methoxypodophyllotoxin [21] which is comparable to the amount of 3.68% described for roots of greenhouse grown plants [22].…”