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

Suresh, B.; Bais, Harsh P.; Raghavarao, K.S.M.S.; Ravishankar, G. A.; Ghildyal, N. P.

doi:10.1016/j.procbio.2003.10.017

Cited by 39 publications

(17 citation statements)

References 23 publications

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“…An inverse relationship between biomass and conductivity was observed not only by our research team, but by Suresh et al [57,58] as well. Authors [57] described this relationship occurred in hairy root culture of Tagetes patula by the same formula as used in P. grandiflorum hairy root shake flask cultures.…”

Section: Discussionsupporting

confidence: 74%

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

et al. 2014

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IntroductionHairy, or transgenic roots are characterized by a rapid and stable growth, which importantly can be initiated and maintained in a hormone-free medium. They often synthetize approximately the same and in some cases even greater amounts of secondary metabolites compared to their mother plants [1,2] and the rate of production remains stable. This makes hairy roots of medicinal plants a good source of valuable compounds and obviously their large-scale culture in bioreactors holds immense potential for the pharmaceutical industry [3].Platycodon grandiflorum (Jacq.) A. DC. (Campanulaceae) is an ornamental plant growing wild in northern Asia, in China, Korea, Japan and east Siberia. For culture it needs light, medium moisture, organically rich, well-drained loams and full sun to part shade.Platycodi radix, the root of P. grandiflorum, has been used as food and in traditional oriental medicine to treat bronchitis, asthma and other pulmonary diseases. Triterpene saponins, called platycosides, are responsible for the medicinal value of this plant material. They have demonstrated a broad spectrum of therapeutic effects, such as antitumor [4][5][6][7], anti-inflammatory [8,9], hepatoprotective [10], antiatherosclerotic [11] and antinociceptive [12]. Additionally, these compounds were found to be a potent adjuvant of specific cellular and humoral immune responses, with the potential use in vaccine production [13][14][15]. Finally, saponins from Platycodi radix stimulated osteoblast differentiation via RUNX2, a principal osteogenic master gene for bone formation [16].Although the extensive pharmacological studies of these valuable compounds were undertaken, to date the sum of saponins, not the chosen, in hairy root cultures of P. grandiflorum was determined only by our research team [17]. Besides, this is the first report of the growth of P. grandiflorum transgenic roots cultured in a mist bioreactor and their yield of saponins. The method of Platycodi radix saponins determination published in "Chinese pharmacopeia" [18] exhibited as no suitable for quantitative analysis of saponins from P. grandiflorum transgenic roots. Therefore, the authors elaborated their own method [17].The compounds mentioned above are the main chemicals found in Platycodi radix. They are typical composed of oleaene backbones with two side chains: one is a 3-O-glucose linked by glycosidic bond, and the other is a 28-O-arabinoserhamnose-xylose-apiose linked by an ester bond [19]. To date, more than 55 triterpenoid saponins have been isolated from Platycodi radix [20]. Examples are platycodins A-I and polygalacins D and D2. According to WHO monographs AbstractThe growth and saponin accumulation were measured in two lines of transgenic hairy roots of Platycodon grandiflorum, Pl 6 and Pl 17, cultured for 8 weeks in 250-ml shake flasks containing 50 ml of hormone-free woody plant medium supplemented with 40 g/l sucrose and in the Pl 17 line cultured for 12 weeks in a 5-l mist bioreactor containing 1.5 l of the same medium. With both methods, the...

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Section: Discussionsupporting

confidence: 74%

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

et al. 2014

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“…Growth was thus expressed as fresh growth index (FGI) (Jacob & Malpathak 2005;Suresh et al 2005). It was expressed as: FGI = (Final fresh weight of biomass -Initial fresh weight of inoculum) / Initial fresh weight of inoculum Isolation and PCR amplification of genomic DNA To determine the presence of Ri T-DNA in hairy root lines, total DNA was extracted from non-transformed roots as a negative control and from hairy roots using the method of Doyle & Doyle (1987) with modifications.…”

Section: Growth Measurementmentioning

confidence: 99%

Induction of Gentiana cruciata hairy roots and their secondary metabolites

et al. 2011

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Gentiana cruciata L. (cross gentian) is a medicinal and ornamental plant. The root extracts of this species are known to exhibit many curative properties. The natural Gentiana populations are exposed to great danger because of their uncontrolled usage. In this study, hairy roots from Gentiana cruciata L. stem and leaf explants belonging to three different clones were induced by inoculation with four different Agrobacterium rhizogenes wild strains namely A4, 15834, 8196 and R1000. Induction of the root transformation was significantly dependent on the explant type used. On the other hand, the genotype and bacterial strain had no significant effect on hairy root formation. Hairy root formation percentages of the explants varied between 5.6-33.3% in the stem explants, and between 0.0-6.7% in the leaf explants. Transformations of the hairy roots were confirmed by PCR using rolC specific primers, and revealed the absence of contaminating A. rhizogenes with virC primers. Total of twelve hairy root clones were obtained, and their secondary metabolite content was also analyzed by HPLC. Quantitative results exhibited that gentiopicroside was the most abundant compound in all root samples. Furthermore, metabolites such as loganic acid, swertiamarin, and sweroside were also identified and quantified in the samples.

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“…Esculin content was almost double as compared to that present in the roots grown in bubble column and nutrient sprinkle bioreactors (Bais et al, 2002). An acoustic mist bioreactor, which enables efficient mass transfer and higher productivity, has been designed by Suresh et al (2005) using Tagetes patula L. hairy roots as a model system.…”

Section: Nutrient Mist Bioreactormentioning

confidence: 99%

Hairy Root Culture for Mass-Production of High-Value Secondary Metabolites

Srivastava

2007

Critical Reviews in Biotechnology

343

158

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Plant cell cultivations are being considered as an alternative to agricultural processes for producing valuable phytochemicals. Since many of these products (secondary metabolites) are obtained by direct extraction from plants grown in natural habitat, several factors can alter their yield. The use of plant cell cultures has overcome several inconveniences for the production of these secondary metabolites. Organized cultures, and especially root cultures, can make a significant contribution in the production of secondary metabolites. Most of the research efforts that use differentiated cultures instead of cell suspension cultures have focused on transformed (hairy) roots. Agrobacterium rhizogenes causes hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection are characterized by high growth rate, genetic stability and growth in hormone free media. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Hairy root cultures offer promise for high production and productivity of valuable secondary metabolites (used as pharmaceuticals, pigments and flavors) in many plants. The main constraint for commercial exploitation of hairy root cultivations is the development and scaling up of appropriate reactor vessels (bioreactors) that permit the growth of interconnected tissues normally unevenly distributed throughout the vessel. Emphasis has focused on designing appropriate bioreactors suitable to culture the delicate and sensitive plant hairy roots. Recent reactors used for mass production of hairy roots can roughly be divided as liquid-phase, gas-phase, or hybrid reactors. The present review highlights the nature, applications, perspectives and scale up of hairy root cultures for the production of valuable secondary metabolites.

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

Cited by 39 publications

References 23 publications

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

Induction of Gentiana cruciata hairy roots and their secondary metabolites

Hairy Root Culture for Mass-Production of High-Value Secondary Metabolites

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