scholarly journals Production of the Anticancer Compound Withaferin A from Genetically Transformed Hairy Root Cultures of Withania Somnifera

2018 ◽  
Vol 13 (8) ◽  
pp. 1934578X1801300
Author(s):  
Zeynab Yousefian ◽  
Behnaz Hosseini ◽  
Hassan Rezadoost ◽  
Javier Palazón ◽  
Mohammad Hossein Mirjalili
Keyword(s):  
Hairy Roots ◽  
Withania Somnifera ◽  
Growth Index ◽  
Dry Weight ◽  
Squalene Synthase ◽  
Withaferin A ◽  
Farnesyl Pyrophosphate ◽  
Transformed Hairy Roots ◽  
Transformed Hairy Root ◽  
Anticancer Compound

Withanolides (WTDs), well-known medicinally important compounds of Withania somnifera, including the anticancer compound withaferin A (WFA), are biosynthesized from their dedicated precursor squalene. Condensation of farnesyl pyrophosphate (FPP) molecules to produce squalene is catalyzed by squalene synthase (SQS). In the present study, the Arabidopsis thaliana squalene synthase gene ( AtSQS1) was transformed in W. somnifera by Agrobacterium tumefaciens C58C1 (pRiA4). The AtSQS1 gene was detected in 87.88 % of induced transformed hairy roots (THRs). The results showed that the growth index (GI) of THRs after five weeks of culture was 2-fold higher than that of adventitious hairy roots (AHRs). The biomass doubling time of THRs and AHRs was 18 and 30 days, respectively. Quantitative evaluation of WFA in the studied roots showed that THRs contain 1.51-fold more WFA (330±0.87μg g−1 dry weight (DW)) than AHRs (218±0.17μg g−1 DW). These findings can be used for the production of WFA as a valuable anticancer compound through controlled and scaled up cultures of W. somnifera THRs.

scholarly journals Xanthones Production in Gentiana dinarica Beck Hairy Root Cultures Grown in Simple Bioreactors

Plants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1610
Author(s):  
Branka Vinterhalter ◽  
Nevena Banjac ◽  
Dragan Vinterhalter ◽  
Dijana Krstić-Milošević
Keyword(s):  
Hairy Root ◽  
Large Scale ◽  
Bubble Column ◽  
Growth Parameters ◽  
Growth Index ◽  
Dry Weight ◽  
Growth Conditions ◽  
Good Growth ◽  
Gentiana Dinarica ◽  
Hairy Root Clones

The hairy root clones of Gentiana dinarica cl-B, cl-D, cl-3, and cl-14 were cultivated in parallel in diverse simple bioreactors, including temporary immersion systems RITA® (TIS RITA®), bubble column bioreactors (BCB), and Erlenmeyer flasks (EF), and evaluated for biomass production and xanthone content. The obtained results showed that TIS RITA® and BCB containing ½ MS medium with 4% sucrose provided equally good growth conditions in which the majority of the clones displayed the higher percentage of dry matter (DM%), and xanthones norswertianin-1-O-primeveroside (nor-1-O-prim) and norswertianin production than those cultivated in EF. Thin and well branched hairy root clone cl-B grown in BCB for 7 weeks was superior regarding all growth parameters tested, including growth index (19.97), dry weight (2.88 g), and DM% (25.70%) compared to all other clones. Cl-B cultured in TIS RITA® contained the highest amount of nor-1-O-prim (56.82 mg per vessel). In BCB with constant aeration, cl-B accumulated the highest norswertianin content reaching 18.08 mg/vessel. The optimized conditions for cultivation of selected G. dinarica hairy root clones in highly aerated TIS RITA® and BCB systems contribute to the development of bioreactor technology designed for the large scale commercial production of xanthones nor-1-O-prim and norswertianin.

scholarly journals Pharmacological Levels of Withaferin A (Withania somnifera) Trigger Clinically Relevant Anticancer Effects Specific to Triple Negative Breast Cancer Cells

PLoS ONE ◽  
2014 ◽  
Vol 9 (2) ◽  
pp. e87850 ◽  
Author(s):  
Katarzyna Szarc vel Szic ◽  
Ken Op de Beeck ◽  
Dariusz Ratman ◽  
An Wouters ◽  
Ilse M. Beck ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Triple Negative Breast Cancer ◽  
Breast Cancer Cells ◽  
Withania Somnifera ◽  
Triple Negative ◽  
Withaferin A ◽  
Anticancer Effects

scholarly journals Reducing Substrate Moisture Content during Greenhouse Production of Poinsettia Improves Postproduction Quality and Economic Value

HortScience ◽  
2018 ◽  
Vol 53 (11) ◽  
pp. 1618-1628
Author(s):  
Yanjun Guo ◽  
Terri Starman ◽  
Charles Hall
Keyword(s):  
Moisture Content ◽  
Economic Value ◽  
Growth Index ◽  
Dry Weight ◽  
Leaf Number ◽  
Stem Length ◽  
Total Production ◽  
Greenhouse Production ◽  
Ethylene Concentration ◽  
Substrate Moisture

The objective was to determine the effect of substrate moisture content (SMC) during poinsettia (Euphorbia pulcherrima) greenhouse production on plant quality, postproduction longevity, and economic value. Two experiments were conducted, one in 2016 with ‘Freedom Red’ and the other in 2017 with ‘Christmas Eve Red’. Treatments included two SMC levels (20% or 40%) applied in four timing of application combinations. Total production (TP) time was 14 (2016) or 12 (2017) weeks in which vegetative production (VP) occurred from week 33 (2016) or 35 (2017) to week 39 and reproductive production (RP) continued from week 40 to 47. The four timing of application treatments were 40/40 = TP at 40% SMC; 20/40 = VP at 20% + RP at 40%; 40/20 = VP at 40% + RP at 20%; 20/20 = TP at 20% SMC. After simulated shipping in the dark, plants were evaluated in a simulated retail environment with two packaging treatments: no sleeve covering or plastic perforated plant sleeves covering container and plant. At the end of greenhouse production, plants grown in 20% SMC during RP (20/20 and 40/20) had shorter bract internode length, stem length, and smaller growth index (GI), decreased shoot and root dry weight (DW), and bract and leaf surface area compared with those in 40% SMC during RP (40/40 and 20/40). Photosynthetic rate was higher when plants were watered at 40% SMC regardless of production stage compared with those in 20% SMC. Leaf thickness, petiole thickness, total bract and leaf number were unaffected by SMC treatments. Plants in 20% SMC during RP (20/20 or 40/20) had earlier bract coloring despite days to anthesis being the same for all SMC treatments. Compared with 40/40, 40/20, and 20/20 could save 44.2% or 43.6%, respectively, irrigation and fertilizer usage, and 39.1% and 47.8%, respectively, labor time. During postharvest, ethylene concentration was unaffected by packaging method. Sleeved plants, regardless of SMC treatment, received lower light intensity in the middle of the plant canopy, causing plants to have lower total leaf number due to abscission and SPAD reading at the end of postproduction. The 40/40 treatment abscised more bracts during five weeks (in 2016) of postproduction and with no sleeve had higher number of bracts with bract edge burn (BEB). In summary, reducing SMC to 20% during TP or RP reduced water usage during production and produced more compact plants with increased postproduction quality.

scholarly journals Effects of Farnesyl Pyrophosphate Accumulation on Calvarial Osteoblast Differentiation

Endocrinology ◽  
2011 ◽  
Vol 152 (8) ◽  
pp. 3113-3122 ◽  
Author(s):  
Megan M. Weivoda ◽  
Raymond J. Hohl
Keyword(s):  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Biosynthetic Pathway ◽  
Squalene Synthase ◽  
Farnesyl Pyrophosphate ◽  
Matrix Mineralization ◽  
Lower Serum Cholesterol ◽  
Calvarial Osteoblast ◽  
Isoprenoid Biosynthetic Pathway

Statins, drugs commonly used to lower serum cholesterol, have been shown to stimulate osteoblast differentiation and bone formation. Statins inhibit 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A reductase (HMGCR), the first step of the isoprenoid biosynthetic pathway, leading to the depletion of the isoprenoids farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP). The effects of statins on bone have previously been attributed to the depletion of GGPP, because the addition of exogenous GGPP prevented statin-stimulated osteoblast differentiation in vitro. However, in a recent report, we demonstrated that the specific depletion of GGPP did not stimulate but, in fact, inhibited osteoblast differentiation. This led us to hypothesize that isoprenoids upstream of GGPP play a role in the regulation of osteoblast differentiation. We demonstrate here that the expression of HMGCR and FPP synthase decreased during primary calvarial osteoblast differentiation, correlating with decreased FPP and GGPP levels during differentiation. Zaragozic acid (ZGA) inhibits the isoprenoid biosynthetic pathway enzyme squalene synthase, leading to an accumulation of the squalene synthase substrate FPP. ZGA treatment of calvarial osteoblasts led to a significant increase in intracellular FPP and resulted in inhibition of osteoblast differentiation as measured by osteoblastic gene expression, alkaline phosphatase activity, and matrix mineralization. Simultaneous HMGCR inhibition prevented the accumulation of FPP and restored osteoblast differentiation. In contrast, specifically inhibiting GGPPS to lower the ZGA-induced increase in GGPP did not restore osteoblast differentiation. The specificity of HMGCR inhibition to restore osteoblast differentiation of ZGA-treated cultures through the reduction in isoprenoid accumulation was confirmed with the addition of exogenous mevalonate. Similar to ZGA treatment, exogenous FPP inhibited the mineralization of primary calvarial osteoblasts. Interestingly, the effects of FPP accumulation on osteoblasts were found to be independent of protein farnesylation. Our findings are the first to demonstrate that the accumulation of FPP impairs osteoblast differentiation and suggests that the depletion of this isoprenoid may be necessary for normal and statin-induced bone formation.

scholarly journals Huperzine A and Huperzine B Production by Prothallus Cultures of Huperzia selago (L.) Bernh. ex Schrank et Mart

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3262
Author(s):  
Wojciech J. Szypuła ◽  
Beata Wileńska ◽  
Aleksandra Misicka ◽  
Agnieszka Pietrosiuk
Keyword(s):  
Pharmaceutical Industry ◽  
Current Source ◽  
Growth Index ◽  
Dry Weight ◽  
Huperzine A ◽  
Biomass Growth ◽  
Tissue Cultures ◽  
In The Wild ◽  
Very High

This is the first report of an efficient and effective procedure to optimize the biosynthesis of huperzine A (HupA) and huperzine B (HupB) in vitro from Huperzia selago gametophytes. Axenic tissue cultures were established using spores collected from the sporophytes growing in the wild. The prothalia were obtained after 7–18 months. Approximately 90 up to 100% of the gametophytes were viable and grew rapidly after each transfer on to a fresh medium every 3 months. The best biomass growth index for prothallus calculated on a fresh (FW) and dry weight (DW) basis, at 24 weeks of culture, was 2500% (FW) and 2200% (DW), respectively. The huperzine A content in the gametophytes was very high and ranged from 0.74 mg/g to 4.73 mg/g DW. The highest yield HupA biosynthesis at >4 mg/g DW was observed on W/S medium without growth regulators at 8 to 24 weeks of culture. The highest HupB content ranged from 0.10 mg/g to 0.52 mg/g DW and was obtained on the same medium. The results demonstrate the superiority of H. selago gametophyte cultures, with the level of HupA biosynthesis approximately 42% higher compared to sporophyte cultures and 35-fold higher than when the alkaloid was isolated from H. serrata, its current source for the pharmaceutical industry. Moreover, the biosynthesis of HupB was several-fold more efficient than in H. selago sporophytes growing in the wild. HPLC-HR-MS analyses of the extracts identified eight new alkaloids previously unreported in H. selago: deacetylfawcettine, fawcettimine, 16-hydroxyhuperzine B, deacetyllycoclavine, annopodine, lycopecurine, des-N-methylfastigiatine and flabelline.

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