Hairy Root Biomass Research Articles

Antioxidant compounds in Salvia officinalis L. shoot and hairy root cultures in the nutrient sprinkle bioreactor

The study focused on the production of compounds with antioxidant activity in hairy root and shoot cultures of <em>Salvia officinalis</em> grown in laboratory-scale sprinkle nutrient bioreactors. HPLC analysis showed that production of rosmarinic acid in transformed roots (34.65 ±1.07 mg l<sup>-1</sup>) was higher that in shoot culture (26.24 ±0.48 mg l<sup>-1</sup>). In the latter diterpenoids: carnosic acid (1.74 ±0.02 mg l<sup>-1</sup>) and carnosol (1.34 ±0.01 mg l<sup>-1</sup>) were also found. Biomass accumulation after a growth period in the bioreactor was also studied. An 18-fold increase in hairy root biomass was recorded after 40 days of culture. In sage shoot culture, biomass increased 43 times after 21 days of bioreactor run. The current operating conditions of the bioreactor were not suitable for the propagation of <em>Salvia officinalis</em> mainly due to the hyperhydricity problem of leaves and stems.

A neural network approach for the prediction of in vitro culture parameters for maximum biomass yields in hairy root cultures

The present study deals with ANN based prediction of culture parameters in terms of inoculum density, pH and volume of growth medium per culture vessel and sucrose content of the growth medium for Glycyrrhiza hairy root cultures. This kind of study could be a model system in exploitation of hairy root cultures for commercial production of pharmaceutical compounds using large bioreactors. The study is aimed to evaluate the efficiency of regression neural network and back propagation neural network for the prediction of optimal culture conditions for maximum hairy root biomass yield. The training data for regression and back propagation networks were primed on the basis of function approximation, where final biomass fresh weight ( f wt ) was considered as a function of culture parameters. On this basis the variables in culture conditions were described in the form of equations which are for inoculum density: y=0.02 x+0.04, for pH of growth medium: y= x+2.8, for sucrose content in medium: y=9.9464 x+(−9.7143) and for culture medium per culture vessel: y=10 x. The fresh weight values obtained from training data were considered as target values and further compared with predicted fresh weight values. The empirical data were used as testing data and further compared with values predicted from trained networks. Standard MATLAB inbuilt generalized regression network with radial basis function radbas as transfer function in layer one and purelin in layer two and back propagation having purelin as transfer function in output layer and logsig in hidden layer were used. Although in comparative assessment both the networks were found efficient for prediction of optimal culture conditions for high biomass production, more accuracy in results was seen with regression network.

Determination of Biomass Composition of Catharanthusroseus Hairy Roots for Metabolic Flux Analysis

Metabolic flux analysis is a powerful diagnostic tool in metabolic engineering, and determination of biomass composition is indispensable to accurate flux evaluation. We report the elemental and biomolecular composition of Catharanthus roseus hairy roots, a pharmaceutically significant plant system and an important metabolic engineering target. The molecular formula of the organic material in the hairy roots was C12.0H22.7N0.4O7.6 during mid-exponential growth. The abundances of lipids, lignin, cellulose, hemicellulose, starch, protein, proteinogenic amino acids, mineral ash, and moisture in the biomass were quantified. Analysis of water-soluble components of the biomass with 1-D 13C and 2-D [1H,1H] correlation (COSY) NMR spectroscopy revealed that the water-soluble components were composed almost entirely of -glucans. Agropine, a frequently reported hairy root biomass component, was not detected. Our measurements of the biomass components quantified 83.6 +/- 9.3% (w/w) of the biomass. Together with previously reported abundances of indole alkaloids, we accounted for at least 85.9 +/- 11.6% (w/w) of the carbon in the biomass, which enabled the precise determination of 12 biomass synthesis fluxes.

Hairy Root Cultures of Gynostemma pentaphyllum (Thunb.) Makino: A Promising Approach for the Production of Gypenosides as an Alternative of Ginseng Saponins

Hairy root cultures of Gynostemma pentaphyllum were established by infecting leaf discs with Agrobacterium rhizogenes. The dry biomass of hairy roots grown in MS medium for 49 days was 7.3 g l(-1) with a gypenoside content of 38 mg g(-1) dry wt.

Comparative evaluation of bioreactor design using Tagetes patula L. hairy roots as a model system

An acoustic mist bioreactor which enables efficient mass transfer and higher productivity has been designed. The design is useful for cultivation of higher density biomass of hairy roots with the added advantage of simplicity in operation. In the present study Tagetes patula hairy root cultures were studied as a model system for the growth and thiophene production in this bioreactor. Results are compared with conventional designs such as bubble column and nutrient sprinkle bioreactors. Hairy roots grown in the acoustic mist bioreactor showed higher biomass (5.4 g DW l −1 after 15 days) and specific growth rate (0.06 per day), comparable with shake flask culture data. The specific growth rate in other designs such as bubble column and nutrient sprinkle was 0.05 and 0.05 per day, respectively. Thiophene content was maximum (2.01 mg l −1 after 15 days) in the acoustic mist bioreactor which was 1.4-fold greater than nutrient sprinkle and bubble column bioreactors and is close to that of shake flask level. The results demonstrate the efficacy and the advantage of the acoustic mist bioreactor for the cultivation of hairy root cultures.

Ginsenoside production by hairy root cultures of Panax ginseng: influence of temperature and light quality

Large-scale production of valuable secondary metabolites by plant cell cultures is generally hampered by low productivity. This productivity is affected by several physical factors like temperature and light. In this study we have investigated the impact of temperature and light quality on biomass accumulation and ginsenoside production by hairy roots cultivated in large-scale bioreactors. Biomass accumulation and ginsenoside production was optimal under 20 °C/13 °C day (12 h)/night (8 h) cycle. Biomass of hairy roots was highest in the cultures grown under dark or red light while ginsenoside accumulation was optimum in the cultures grown under fluorescent light.

Growth and betacyanin production by hairy roots of Beta vulgaris in airlift bioreactors

Hairy roots of red beet (Beta vulgaris L.) were cultivated in different types of airlift bioreactors (cone, balloon, bulb, drum and column bioreactors of 5 l capacity and containing 3 l of half strength Murashige & Skoog medium). The cone type of airlift bioreactor gave the highest biomass of hairy roots and betacyanin accumulation. Betacyanin accumulation was 27 mg g−1 dry wt in cultures aerated at 0.3 vvm. Light irradiation of 20 μmol m−2 s−1 promoted hairy root growth but optimum betacyanin (34 mg g−1 dry wt) accumulation was with the cultures grown under 60 μmol m−2 s−1.

Improved growth of Artemisia annua L hairy roots and artemisinin production under red light conditions

Hairy root cultures of Artemisia annua L were cultivated for 30 days under either white, red, blue, yellow or green light. Red light at 660 nm gave the highest biomass of hairy roots (5.73 g dry wt cells l−1 medium) and artemisinin content (31 mg arteminsinin g−1 dry cells) which were, respectively, 17% and 67% higher than those obtained under white light.