Journal articles: 'Hairi root cultures'

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Yeo, Hyeon Ji, Min Jae Kwon, Sang Yeon Han, Jae Cheol Jeong, Cha Young Kim, Sang Un Park, and Chang Ha Park. "Effects of Carbohydrates on Rosmarinic Acid Production and In Vitro Antimicrobial Activities in Hairy Root Cultures of Agastache rugosa." Plants 12, no. 4 (February 10, 2023): 797. http://dx.doi.org/10.3390/plants12040797.

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Agastache rugosa (popularly known as Korean mint) belongs to the Lamiaceae family and comprises 22 species of perennial aromatic medicinal species native to East Asian countries, such as Korea, Taiwan, Japan, and China. A. rugosa contains many phenolic compounds that exhibit pharmacological and physiological activities, including antioxidant, anticancer, antiviral, antifungal, and antibacterial activities. The highest concentrations of rosmarinic acid and its isomers have been reported in the roots of A. rugosa. In this in vitro study, hairy roots of A. rugosa were obtained and the carbohydrates (sorbitol, mannitol, glucose, maltose, galactose, mannose, and sucrose) were evaluated to determine those that were optimal for rosmarinic acid production and hairy root growth. Antioxidant and antibacterial activities of extracts of A. rugosa were also assessed. The best carbon source for A. rugosa hairy root cultures was sucrose, considering biomass productivity (0.460 ± 0.034 mg/30 mL), rosmarinic acid production (7.656 ± 0.407 mg/g dry weight), and total phenolic content (12.714 ± 0.202 mg/g gallic acid equivalent). Antioxidant and antimicrobial activities were displayed by A. rugosa hairy roots cultured in liquid medium supplemented with 100 mM sucrose. Twenty-five bacterial strains, including multidrug-resistant bacteria and one pathogenic yeast strain, were used for antimicrobial screening of A. rugosa hairy roots. The hairy root extracts displayed antibacterial activity against Micrococcus luteus (KCTC 3063) and Bacillus cereus (KCTC 3624). The inhibition of these bacteria was greater using A. rugosa hairy roots with the highest levels of phenolic compounds cultured in the presence of sucrose, compared to hairy roots with the lowest levels of phenolic compounds cultured in the presence of fructose. Considering hairy root biomass, phenolic compound production, and antibacterial activity, sucrose is the best carbon source for A. rugosa hairy root cultures.

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Drobot, K. O. "TARRAGON (Artemisia dracunculus L.) “HAIRY” ROOT CULTURE PRODUCTION." Biotechnologia Acta 9, no. 2 (2016): 55–60. http://dx.doi.org/10.15407/biotech9.02.055.

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Mohagheghzadeh, Abdolali, Azra Gholami, Shiva Hemmati, and Shadab Dehshahri. "Bag Culture: A Method for Root-Root Co-Culture." Zeitschrift für Naturforschung C 63, no. 1-2 (February 1, 2008): 157–60. http://dx.doi.org/10.1515/znc-2008-1-229.

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A method named “bag culture” was developed for coculturing of Linum persicum (section Syllinum) and L. austriacum (section Linum) hairy roots. For this propose L. austriacum and L. persicum hairy root cultures were established using Agrobacterium rhizogenes in McCown medium. L. persicum hairy roots in bags (1 mm2 mesh) were successfully grown together with L. austriacum hairy roots. The amounts of podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (MPTOX) produced by L. persicum hairy root cultures were detected using HPLC. The results indicated that the amounts of both lignans and growth indexes of the two hairy roots decreased, that may be partly due to a competition between the two types of culture in using precursors of biosynthetic metabolites and the amount of culture medium which is available for each hairy root. However, MPTOX (0.17 g/100 g DW) and PTOX (0.02 g/100 g DW) levels of the L. persicum single culture in bag were significantly higher than of the other cultures which may be due to the immobilization effect of the bag.

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D'Angiolillo, Francesca, Cecilia Noccioli, Barbara Ruffoni, Roberto Scarpato, Luisa Pistelli, and Laura Pistelli. "Daidzein Production and HeLa Cytotoxicity of Bituminaria bituminosa Hairy Root Cultures." Natural Product Communications 12, no. 11 (November 2017): 1934578X1701201. http://dx.doi.org/10.1177/1934578x1701201119.

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Bituminaria bituminosa (L.) C.H. Stirt is a perennial species widely distributed in the Mediterranean basin and the Canary Islands. This species is used in folk medicine and currently has considerable pharmaceutical interest for its content in phenylpropanoids, furanocoumarins and pterocarpans. In vitro cultures (shoots and hairy roots) have been performed to obtain plant material useful for the production of these metabolites. Hairy root cultures were successfully established after inoculation of hypocotyls with the LBA 9402 A. rhizogenes strain. The HRPB3 line was selected for further analysis and elicited with chitosan and salicylic acid. All the HRPB3 cultures showed higher polyphenol content and greater DPPH-antioxidant activity than shoots cultured in vitro. The presence of isoflavone daidzein was detected in the hairy root extracts. The cytotoxic effect of HR extracts has been further tested on HeLa cells: the salicylic acid elicited HR exhibited good antiproliferative effects.

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Kuzovkina, I. N., A. Gohar, and I. E. Alterman. "Production of β-Carboline Alkaloids in Transformed Root Cultures of Peganum harmala L." Zeitschrift für Naturforschung C 45, no. 6 (June 1, 1990): 727–28. http://dx.doi.org/10.1515/znc-1990-0626.

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Abstract Peganum harmala, Hairy Root Cultures, β-Carboline Alkaloids Hairy root cultures of Peganum harmala were established by genetic transformation with Agrobacterium rhizogenes strain A4. In contrast to suspension cultures the root cultures contained high levels of β-carboline alka loids (1 -1.5% of dry mass).

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C, Veeresham, C. S. Reddy, and Praveena Ch. "Strategies to Improve the Production of Forskolin from Hairy Root Cultures of Coleus forskohlii Briq." International Journal of Pharmaceutical Sciences and Nanotechnology 5, no. 2 (August 31, 2012): 1720–26. http://dx.doi.org/10.37285/ijpsn.2012.5.2.7.

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The aim of this study was to elucidate the effect of elicitors and precursors on the production of forskolin from the hairy root cultures of Coleus forskohlii Briq. Hairy root cultures were established from leaf explants by infecting with Agrobacterium rhizogenes strain A4 on MS basal medium. Suspension cultures of hairy root cultures were initiated in MS medium containing IBA (1.0 mg/L), casein hydrolysate (600 mg/L). We investigated the growth of biomass and forskolin production in suspension cultures of hairy roots. The production of forskolin was parallel to the growth of biomass. The maximum production of forskolin was observed after 5 weeks. With the objective to increase the yield of forskolin, abiotic elicitors such as salicylic acid (100 μM and 500 μM), copper sulphate (100 μM and 500 μM), methyl jasmonate (100 μM and 500 μM) and precursors such as α-ketoglutaric acid (0.2 mM and 1.0 mM), L-phenylalanine (0.2 mM and 1.0 mM) were added to hairy root cultures on different days of incubation period and evaluated their effects on production of forskolin. Elicitor, methyl jasmonate (500 μM) and the precursor, L-phenylalanine (1 mM) on day-14 addition significantly enhanced the production of forskolin over the control hairy root cultures C. forskohlii. Given forskolin’s limited commercial supply, this study provides avenues for improving the production of forskolin in the hairy root culture of C. forskohlii.

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Jeziorek, Małgorzata, Katarzyna Sykłowska-Baranek, and Agnieszka Pietrosiuk. "Hairy Root Cultures for the Production of Anti-cancer Naphthoquinone Compounds." Current Medicinal Chemistry 25, no. 36 (December 3, 2018): 4718–39. http://dx.doi.org/10.2174/0929867324666170821161844.

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Background: Recent years have brought the dynamic development in studies of naphthoquinones obtained from plants, in vitro cultures and semi- or total synthesis. This review presents the hairy root cultures approach for producing naphthoquinones and summarizes their most recent anti-cancer investigations. Objective: This review aimed to define biotechnological strategies impacted on naphthoquinones production in hairy root cultures. Up to now the major source of shikonin/alkannin derivatives, rhinacanthins and ramentaceone is isolation from plant material, also derived via biotechnological methods. Moreover, the most recent anti-cancer activity studies on naphthoquinones which could be produced in hairy root cultures were outlined. Methods: For databases survey two selection criteria were used: (i) naphthoquinone could be produced in hairy roots, and (ii) it exhibits anti-cancer properties. Results: Ninety two papers were included in the review, thirty described biotechnological approaches enhancing naphthoquinones production, among them twenty seven were dedicated to hairy root cultures. Forty papers outlined the anti-cancer activity of targeted naphthoquinones including the type of cancer and bioassays description. The synergistic effect of natural naphthoquinones and other anti-cancer therapies was reviewed and toxicity of natural naphthoquinones and plant extracts was discussed. The review highlights tendencies in hairy root investigations and indicates the possible future research directions for improving biotechnological production efficacy. Conclusion: This review demonstrates a great potential of hairy root cultures for naphthoquinones production, which could be furtherly developed for future medical purposes, especially as anti-cancer agents. This area of plant biotechnology will be surely still developed with traditional and new strategies.

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Berkov, Strahil, Atanas Pavlov, Petia Kovatcheva, Pepa Stanimirova, and Stefan Philipov. "Alkaloid Spectrum in Diploid and Tetraploid Hairy Root Cultures of Datura stramonium." Zeitschrift für Naturforschung C 58, no. 1-2 (February 1, 2003): 42–46. http://dx.doi.org/10.1515/znc-2003-1-207.

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Hairy root cultures were obtained from diploid and induced tetraploid plants of Datura stramonium and analyzed by gas chromatography/mass spectrometry. Twenty alkaloids (19 for diploid and 9 for tetraploid hairy root cultures) were identified. A new tropane ester 3-tigloyloxy-6-propionyloxy-7-hydroxytropane was identified on the basis of mass spectral data. Hyoscyamine was the main alkaloid in both diploid and tetraploid cultures. In contrast to diploid hairy roots, the percentage contributions of the alkaloids, with exceptions for hyoscyamine and apoatropine, were higher in the total alkaloid mixture of tetraploid hairy roots

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Wysokińska, Halina, Katarzyna Lisowska, and Katarzyna Floryanowicz-Czekalska. "Transformation of Catalpa ovata by Agrobacterium rhizogenes and Phenylethanoid Glycosides Production in Transformed Root Cultures." Zeitschrift für Naturforschung C 56, no. 5-6 (June 1, 2001): 375–81. http://dx.doi.org/10.1515/znc-2001-5-610.

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Transformed root cultures of Catalpa ovata were established following shoots infection with four agropine strains of Agrobacterium rhizogenes. Frequency of root formation was dependent on the bacterial strain and the presence of acetosyringone in the incubation medium. It is the first report concerning the possibility of transforming Catalpa ovata by A. rhizogenes. Both transformed and untransformed root cultures of C. ovata were studied for their growth and phenylethanoid glycoside production. As with the roots of intact plants, cis-and trans-verbascoside as well as martynoside were produced in transformed and untransformed root cultures of C. ovata. In hairy roots, total (cis + trans) verbascoside production could be stimulated up to three-fold of that of roots of 6-month-old plants grown in a greenhouse, by using an appropriate root line cultured in liquid 1/2 B5 Gamborg medium containing indole-3-butyric acid (0.1 mg/1) in the dark but not light conditions. Transformed and untransformed root cultures of C. ovata were also found to have 10 times higher martynoside production than roots of intact plants.

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Kim, Sun-Ju, Woo Tae Park, Md Romij Uddin, Yeon Bok Kim, Sang-Yong Nam, Kwang Hyun Jho, and Sang Un Park. "Glucosinolate Biosynthesis in Hairy Root Cultures of Broccoli (Brassica oleracea var. italica)." Natural Product Communications 8, no. 2 (February 2013): 1934578X1300800. http://dx.doi.org/10.1177/1934578x1300800222.

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Here we present previously unreported glucosinolate production by hairy root cultures of broccoli (B. oleracea var. italica). Growth media greatly influenced the growth and glucosinolate content of hairy root cultures of broccoli. Seven glucosinolates, glucoraphanin, gluconapin, glucoerucin, glucobrassicin, 4- methoxyglucobrassicin, gluconasturtiin, and neoglucobrassicin, were identified by analysis of the broccoli hairy root cultures. Both half and full strength B5 and SH media enabled the highest accumulation of glucosinolates. In most cases, the levels of glucosinolates were higher in SH and BS media. Among the 7 glucosinolates, the accumulation of neoglucobrassicin was very high, irrespective of growth medium. The neoglucobrassicin content was 7.4-fold higher in SH medium than 1/2 MS, in which its level was the lowest. The 1/2 B5 medium supported the production of the highest amounts of glucobrassicin and 4- methoxyglucobrassicin, the levels for which were 36.2- and 7.9- fold higher, respectively, than their lowest content in 1/2 MS medium. The 1/2 SH medium enabled the highest accumulation of glucoraphanin and gluconapin in the broccoli hairy root cultures, whose levels were 1.8- and 4.6-fold higher, respectively, than their lowest content in 1/2 MS medium. Our results suggest that hairy root cultures of broccoli could be a valuable alternative approach for the production of glucosinolate compounds.

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Vojin, Tadić, Milošević Snežana, Cingel Aleksandar, Petrić Marija, Trifunović Milana, Antonić Dragana, Tadić Jovan, and Subotić Angelina. "Production of hairy root cultures of lettuce (Lactuca sativa L.)." Open Life Sciences 9, no. 12 (December 1, 2014): 1196–205. http://dx.doi.org/10.2478/s11535-014-0351-9.

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AbstractHairy root cultures of lettuce (Lactuca sativa L.) were obtained by inoculation of cotyledonary leaves of in vitro lettuce seedlings (cvs. Nansen and Ljubljanska ledenka) with Agrobacterium rhizogenes A4M70GUS. Approximately in 96.7% cvs. Nansen and in 91.2% Ljubljanska ledenka inoculated explants produced hairy root when they were incubated on Murashige and Skoog (MS) half-strength medium without plant growth regulators. A total of 54% of all hairy root cultures expressed GUS activity. Every hairy root represented an independent transformation event. Line Ljubljanska ledenka 18 showed the highest biomass (5.5 times the biomass of control root). A PCR analysis of the genomic DNA confirmed the presence of marker and target genes in 15 hairy roots examined.

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Hussain, Masooma Jawad, Yawar Abbas, Naushaba Nazli, Sara Fatima, Samantha Drouet, Christophe Hano, and Bilal Haider Abbasi. "Root Cultures, a Boon for the Production of Valuable Compounds: A Comparative Review." Plants 11, no. 3 (February 5, 2022): 439. http://dx.doi.org/10.3390/plants11030439.

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Medicinal plants are an inevitable source of pharmaceutical drugs and most of the world population depends on these plants for health benefits. The increasing global demand for bioactive compounds from medicinal plants has posed a great threat to their existence due to overexploitation. Adventitious root and hairy root culture systems are an alternative approach to the conventional method for mass production of valuable compounds from medicinal plants owing to their rapid growth, biosynthetic and genetic stability. The main purpose of this review is to investigate the recent scientific research published worldwide on the application of adventitious and hairy root cultures to produce valuable compounds from medicinal plants. Furthermore, a comparison of adventitious root vs. hairy root cultures to produce valuable compounds has also been discussed. Various aspects such as medium composition, carbon source, pH, amount of macronutrients, optimization strategy, scale-up cultures, and use of biotic abiotic and nano-elicitors at various concentrations are the topic of discussion in this review. Several studies on adventitious and hairy root cultures of Polygonum multiflorum¸ Withania somnifera¸ Echinacea purpurea and Ajuga bracteosa have been discussed in detail which highlights the importance of elicitation strategies and bioreactor system, presenting commercial applications.

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Urbańska, Natalia, Joanna Giebułtowicz, Olga Olszowska, and Wojciech J. Szypuła. "The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor." Acta Societatis Botanicorum Poloniae 83, no. 3 (2014): 229–37. http://dx.doi.org/10.5586/asbp.2014.017.

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The 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 growth of transgenic hairy roots was assessed as both fresh and dry weight and the biomass growth was correlated with the conductivity and sucrose uptake. The accumulation of saponins was measured and compared with that in roots derived from the field cultivation. The saponin concentrations were significantly higher in the two hairy root lines cultured in shake flasks [6.92 g/100 g d.w. (g%) and 5.82 g% in Pl 6 and Pl 17, respectively] and the line cultured in the bioreactor (5.93 g%) than in the roots derived from the field cultivation (4.02 g%). The results suggest that cultures of P. grandiflorum hairy roots may be a valuable source for obtaining saponins.

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ZHAO, Shuang, and Hong TANG. "Enhanced production of valtrate in hairy root cultures of Valeriana jatamansi Jones by methyl jasmonate, jasmonic acid and salicylic acid elicitors." Notulae Botanicae Horti Agrobotanici Cluj-Napoca 48, no. 2 (June 30, 2020): 839–48. http://dx.doi.org/10.15835/nbha48211891.

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Valtrate is a pharmacologically active epoxyiridoid ester found in the roots and rhizomes of Valeriana jatamansi Jones. The plant produces only small amounts of this metabolite naturally, and so induction of hairy roots as well as elicitation can be useful to increase its commercial production. In this study, strain R1601 of Agrobacterium rhizogenes was used to induce hairy roots in V. jatamansi, and stable hairy root cultures of V. jatamansi were established successfully. The influence of three exogenous elicitors including methyl jasmonate (MJ), jasmonic acid (JA) and salicylic acid (SA) on valtrate production in the hairy root cultures of V. jatamansi was also investigated, and the 25-day-old hairy root cultures were treated with different concentrations of the elicitors at exposure time of 7 days. This present study showed that MJ (100 mg/L) highly promoted valtrate production at 7 days after elicitation, to a level of 3.63 times higher than that of non-elicited control. SA did not significantly increase the production of valtrate. This is the first-time study to assess the elicitation of hairy root cultures to promote valtrate biosynthesis in V. jatamansi and the resulting experiments demonstrated that MJ was indeed a potent inducer of valtrate biosynthesis.

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Bashan, Yoav, and Hanna Levanony. "Factors affecting adsorption of Azospirillum brasilense Cd to root hairs as compared with root surface of wheat." Canadian Journal of Microbiology 35, no. 10 (October 1, 1989): 936–44. http://dx.doi.org/10.1139/m89-155.

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Electron microscopy of wheat (Triticum aestivum) roots inoculated with Azospirillum brasilense Cd revealed massive adsorption of bacterial cells to the root surface and less adsorption to root hairs. Quantitative analysis of A. brasilense Cd adsorption to root surface and to root hairs, confirmed qualitatively by light microscopy observations, revealed a bacterial adsorption ratio of 5 (+2): 1 (root surface: root hairs). Extreme bacterial adsorption ratios were recorded when bacteria were previously grown in the presence of KNO3 (27:1) or when bacterial cells were inoculated under hydroponic plant growth conditions (80:1). Adsorption of A. brasilense Cd to roots was directly related to the bacterial growth phase, with logarithmic phase cultures demonstrating a greater adsorption than stationary phase cultures. Adsorption to root hairs was dependent mainly on the number of root hairs developed under certain growth conditions. When very few root hairs had developed, most of the bacterial cells were adsorbed to the root surface. Factors such as starvation, bacteria grown in culture in the presence of KNO3, addition of several nutrients, and protease or NaEDTA treatments of bacterial cells before the adsorption assay decreased bacterial adsorption to root hairs. Other factors such as microaerophilic growth conditions, addition of several bacterial chemoattractants, and cellulase-treated root hairs enhanced bacterial adsorption. It is proposed that although A. brasilense Cd adsorbed to every part of the root system, more cells adsorbed to the root surface of wheat than to the root hairs.Key words: associative bacteria, Azospirillum, bacterial adsorption, beneficial bacteria, rhizosphere bacteria, root-hair colonization.

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Walters, K. S., R. D. Sjolund, and K. C. Moore. "SEM and fluorescence imaging of callose deposition in root hair tips and suspension cultures of Streptanthus tortuosus." Proceedings, annual meeting, Electron Microscopy Society of America 48, no. 3 (August 12, 1990): 698–99. http://dx.doi.org/10.1017/s0424820100161047.

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Callose, B-1,3-glucan, a component of cell walls, is associated with phloem sieve plates, plasmodesmata, and other cell wall structures that are formed in response to wounding or infection. Callose reacts with aniline blue to form a fluorescent complex that can be recognized in the light microscope with ultraviolet illumination. We have identified callose in cell wall protuberances that are formed spontaneously in suspension-cultured cells of S. tortuosus and in the tips of root hairs formed in sterile callus cultures of S. tortuosus. Callose deposits in root hairs are restricted to root hair tips which appear to be damaged or deformed, while normal root hair tips lack callose deposits. The callose deposits found in suspension culture cells are restricted to regions where unusual outgrowths or protuberances are formed on the cell surfaces, specifically regions that are the sites of new cell wall formation.Callose formation has been shown to be regulated by intracellular calcium levels.

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Kochan, Ewa, Adriana Nowak, Małgorzata Zakłos-Szyda, Daria Szczuka, Grażyna Szymańska, and Ilona Motyl. "Panax quinquefolium L. Ginsenosides from Hairy Root Cultures and Their Clones Exert Cytotoxic, Genotoxic and Pro-Apoptotic Activity towards Human Colon Adenocarcinoma Cell Line Caco-2." Molecules 25, no. 9 (May 11, 2020): 2262. http://dx.doi.org/10.3390/molecules25092262.

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American ginseng, Panax quinquefolium (L.), is traditionally used in folk medicine. It exhibits a range of anti-inflammatory, hepatoprotective, anti-diabetic, anti-obesity, anti-hyperlipidemic and anti-carcinogenic effects. Its main components are ginsenosides, also known as panaxosides or triterpene saponins. In order to obtain high yields of ginsenosides, different methods of controlled production are involved, i.e., with hairy root cultures. However, they are still employed under in vitro conditions. Our studies revealed that hairy root cultures subjected to an elicitation process can be considered as a potent source of ginsenosides. The present study examines the biological activity of ginseng hairy root cultures against the Caco-2 human adenocarcinoma cell line. Among our six different clones of P. quinquefolium hairy roots, extracts B and Be (treated with elicitor) were the strongest inhibitors of the cellular metabolic activity. While all extracts induced DNA damage, B and Be also generated reactive oxygen species (ROS) in a concentration-dependent manner, which was correlated with the depletion of the mitochondrial membrane potential and induction of apoptosis. These findings indicate that further research concerning P. quinquefolium hairy root cultures should focus on the activity of rare ginsenosides and other biologically active compound profiles (i.e., phenolic compounds).

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Wink, Michael, A. Wilhelm Alfermann, Rochus Franke, Bernhard Wetterauer, Melanie Distl, Jörg Windhövel, Oliver Krohn, et al. "Sustainable bioproduction of phytochemicals by plant in vitro cultures: anticancer agents." Plant Genetic Resources 3, no. 2 (August 2005): 90–100. http://dx.doi.org/10.1079/pgr200575.

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Due to their complex structure with several chiral centres important anticancer agents are still extracted from plants and not synthesized chemically on a commercial scale. Sustainable bioproduction of the compounds of interest may be achieved by plant in vitro cultures. Undifferentiated callus and suspension cultures, which can be cultivated in large bioreactors easily, very often fail to accumulate the compounds of interest, whereas shoot and root cultures as well hairy roots normally produce the same compounds as in the appropriate organs. The production of anticancer compounds, such as the alkaloids vinblastine, vincristine, paclitaxel (Taxol®), camptothecin, or the lignan podophyllotoxin, by plant in vitro cultures is reviewed. Taxanes can be produced in bioreactors using cell suspensions of various Taxus species with good yields; presently paclitaxel is produced on a commercial scale by Phyton Biotech (Germany). Camptothecin has low yields in suspension cultures of Camptotheca acuminata or Nothapodytes foetida (0.0003–0.01%), but a good production (0.1–0.3% dry wt) in root and hairy root cultures of Ophiorrhiza pumila, O. mungos and C. acuminata. Podophyllotoxin can be produced in cell suspension and root as well as hairy root cultures of Podophyllum and various Linum species up to 130 mg/l (Linum album cell suspensions); its derivative 6-methoxypodophyllotoxin is accumulated in hairy roots of L. persicum up to about 500 mg/l. The in vitro production of dimeric indole alkaloids in Catharanthus roseus has failed so far both in undifferentiated and differentiated in vitro cultures. In cases where in vitro cultures show good yields, they can be employed in biotechnology for the sustainable production of valuable products.

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Srinivasan, M., K. Kumar, K. Kumutha, and P. Marimuthu. "Establishing monoxenic culture of arbuscular mycorrhizal fungus Glomus intraradices through root organ culture." Journal of Applied and Natural Science 6, no. 1 (June 1, 2014): 290–93. http://dx.doi.org/10.31018/jans.v6i1.417.

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Arbuscular mycorrhizal fungi are soil fungi distributed worldwide, forming symbiosis with most of the vascular plants for their growth and survival, which is used for sustainable agriculture and ecosystem management. This study investigated the establishment of monoxenic cultures of Glomus intraradices in association with transformed carrot hairy root. The G.intraradices spores were isolated from sugarcane rhizosphere by wet sieving and decanting technique and propagated in open pot culture. Transformation in to carrot hairy root was done using Agrobacterium rhizogenes. Surface sterilization of G.intraradices spores co-cultured with transformed carrot hairy root in Modified Strulla and Romand (MSR) medium was found the host root growth as well as for germination AM spores. After three months of incubation in dark condition, significant production of extensive hyphal growth on MSR medium and an average of 8500-9000 spores per petri dish was observed. The in vitro inoculum exhibited higher potential of root colonization due to numerous intraradices mycelium with extensive spore load. The produced monoxenic inoculum can be used in place of traditional system where it has a advantage of producing contaminant free propagulas. Thus the monoxenic culture system, a powerful tool, of AM sporulation, can be used for the mass production of monoxenic inoculum of AM fungi besides studying its biology.

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Kim, Yeon Bok, Darwin W. Reed, and Patrick S. Covello. "Production of Triterpenoid Sapogenins in Hairy Root Cultures of Silene vulgaris." Natural Product Communications 10, no. 11 (November 2015): 1934578X1501001. http://dx.doi.org/10.1177/1934578x1501001129.

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Silene vulgaris (Moench) Garcke (Caryophyllaceae) is widely distributed in North America and contains bioactive oleanane-type saponins. In order to investigate in vitro production of triterpenoid saponins, hairy root cultures of S. vulgaris were established by infecting leaf explants with five strains of Agrobacterium rhizogenes (LBA9402, R1000, A4, 13333, and 15834). The A. rhizogenes strain LBA9402 had an infection of 100% frequency and induced the most hairy roots per plant. Methyl jasmonate (MeJA)-induced changes in triterpenoid saponins in S. vulgaris hairy roots were analyzed. Accumulation of segetalic acid and gypsogenic acid after MeJA treatment was 5-and 2-fold higher, respectively, than that of control root. We suggest that hairy root cultures of S. vulgaris could be an important alternative approach to the production of saponins.

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Berlin, J., L. Fecker, C. Rügenhagen, C. Sator, D. Strack, L. Witte, and V. Wray. "Isoflavone Glycoside Formation in Transformed and Non-Transformed Suspension and Hairy Root Cultures of Lupinus polyphyllus and Lupinus hartwegii." Zeitschrift für Naturforschung C 46, no. 9-10 (October 1, 1991): 725–34. http://dx.doi.org/10.1515/znc-1991-9-1003.

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Abstract Transformed cell suspension and hairy root cultures were established by infecting seedlings of Lupinus polyphyllus and L. hartwegii with various wild type strains of Agrobacterium tume-faciens and A. rhizogenes. Transformation of the cultures was confirmed either by their phytohormone autotrophy, detection of opines or southern analysis. Glueosides of genistein and 2′-hydroxygenistein, were found to be the main secondary metabolites in normal and transformed suspension cultures as well as in hairy root cultures. Although some of the isoflavone glycosides of the cultures were apparently new constituents of Lupinus, they were afterwards also found in young seedlings.

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Komarovská, Hedviga, Annalisa Giovannini, Ján Košuth, and Eva Čellárová. "Agrobacterium rhizogenes-Mediated Transformation of Hypericum tomentosum L. and Hypericum tetrapterum Fries." Zeitschrift für Naturforschung C 64, no. 11-12 (December 1, 2009): 864–68. http://dx.doi.org/10.1515/znc-2009-11-1218.

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This is the fi rst evidence on successful Agrobacterium rhizogenes-mediated genetic transformation of two species from the genus Hypericum, H. tomentosum L. and H. tetrapterum Fries. Hairy root cultures were induced from root segments of both Hypericum species by two agropine wild-type strains of A. rhizogenes, ATCC 15834 and A4. The transgenic character of the hairy root cultures was proved by PCR amplifi cation of the rolABCD genes. In some H. tetrapterum transgenic lines aux genes were detected as well.

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23

SHANKS, J., and J. MORGAN. "Plant ‘hairy root’ culture." Current Opinion in Biotechnology 10, no. 2 (April 1, 1999): 151–55. http://dx.doi.org/10.1016/s0958-1669(99)80026-3.

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24

YS, Wang, Jensen LS, and J. Magid. "Differential responses of root and root hair traits of spring wheat genotypes to phosphorus deficiency in solution culture." Plant, Soil and Environment 62, No. 12 (November 24, 2016): 540–46. http://dx.doi.org/10.17221/485/2016-pse.

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25

Sharma, Amit Raj, Gaurav Gajurel, Izzeldin Ahmed, Krystian Roedel, and Fabricio Medina-Bolivar. "Induction of the Prenylated Stilbenoids Arachidin-1 and Arachidin-3 and Their Semi-Preparative Separation and Purification from Hairy Root Cultures of Peanut (Arachis hypogaea L.)." Molecules 27, no. 18 (September 19, 2022): 6118. http://dx.doi.org/10.3390/molecules27186118.

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Prenylated stilbenoids such as arachidin-1 and arachidin-3 are stilbene derivatives that exhibit multiple pharmacological activities. We report an elicitation strategy using different combinations of cyclodextrin, hydrogen peroxide, methyl jasmonate and magnesium chloride to increase arachidin-1 and arachidin-3 production in peanut hairy root cultures. The treatment of hairy root cultures with cyclodextrin with hydrogen peroxide selectively enhanced arachidin-1 yield (132.6 ± 20.4 mg/L), which was 1.8-fold higher than arachidin-3. Similarly, cyclodextrin combined with methyl jasmonate selectively enhanced arachidin-3 yield (178.2 ± 6.8 mg/L), which was 5.5-fold higher than arachidin-1. Re-elicitation of the hairy root cultures further increased the levels of arachidin-1 and arachidin-3 by 24% and 42%, respectively. The ethyl acetate extract of the culture medium was consecutively fractionated by normal- and reversed-phase column chromatography, followed by semi-preparative HPLC purification on a C18 column to yield arachidin-1 with a recovery rate of 32% and arachidin-3 with a recovery rate of 39%, both at higher than 95% purity. This study provided a sustainable strategy to produce high-purity arachidin-1 and arachidin-3 using hairy root cultures of peanuts combined with column chromatography and semi-preparative HPLC.

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26

Lee, Sang-Won, Young Seon Kim, Md Romij Uddin, Do Yeon Kwon, Yeon Bok Kim, Mi Young Lee, Sun-Ju Kim, and Sang Un Park. "Resveratrol Production from Hairy Root Cultures of Scutellaria baicalensis." Natural Product Communications 8, no. 5 (May 2013): 1934578X1300800. http://dx.doi.org/10.1177/1934578x1300800517.

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The levels of resveratrol produced by hairy root cultures of Scutellaria baicalensis were investigated using different media of varying strengths and in the presence of various concentrations of auxins. The levels of resveratrol were higher when the hairy root cultures were maintained in full-strength Murashige and Skoog (MS) medium when compared with other growth media. The cultures grown in full-strength MS medium produced 2.5-fold higher resveratrol than those grown in half-strength B5 medium—the lowest resveratrol-producing medium. The levels of resveratrol varied significantly when cultures were grown in full-strength MS with varying concentrations of auxins. Supplementation of the media with the auxin indole acetic acid (IAA) at 0.1 mg/L produced the highest accumulation of resveratrol. Our findings reveal a valuable alternative approach for the production of resveratrol from S. baicalensis.

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27

Fang, Lingling, Amit Raj Sharma, Chineche Aniemena, Krystian Roedel, Florence Henry, Philippe Moussou, Anita Samuga, and Fabricio Medina-Bolivar. "Elicitation of Stilbenes and Benzofuran Derivatives in Hairy Root Cultures of White Mulberry (Morus alba)." Plants 12, no. 1 (December 31, 2022): 175. http://dx.doi.org/10.3390/plants12010175.

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Stilbene and benzofuran derivatives isolated from the root of white mulberry (Morus alba) have shown various biological activities, including anti-inflammatory, antioxidant, and antimicrobial properties. The objectives of this study were to develop hairy root cultures and assess the effect of multiple elicitors combinations including (I) methyl-β-cyclodextrin (CD), MgCl2, methyl jasmonate (MeJA), and H2O2, (II) CD, MgCl2, and MeJA and (III) CD, MgCl2, and H2O2, on the production of these bioactive compounds. The highest yields of stilbenes and benzofurans were obtained upon co-treatment with 18 g/L CD, 3 mM H2O2 and 1 mM MgCl2. The stilbenes oxyresveratrol, resveratrol, and 3′-prenylresveratrol accumulated up to 6.27, 0.61, and 5.00 mg/g DW root, respectively. Meanwhile, the aryl benzofurans moracin M and moracin C accumulated up to 7.82 and 1.82 mg/g DW root, respectively. These stilbenes and benzofurans accumulated in the culture medium of the elicited hairy root cultures. They were not detected in the root tissue. However, the oxyresveratrol diglucoside mulberroside A was only detected in the root tissue with yields up to 10.01 mg/g DW. The results demonstrated that co-treatment of white mulberry hairy root cultures with multiple elicitors can significantly enhance production and secretion of stilbenes and benzofurans in this controlled and sustainable axenic culture system.

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28

Naicker, Leeann, Viresh Mohanlall, and Bharti Odhav. "GENETIC TRANSFORMATION OF CERATOTHECA TRILOBA FOR THE PRODUCTION OF ANTHRAQUINONES FROM HAIRY ROOT CULTURES." African Journal of Traditional, Complementary and Alternative Medicines 13, no. 3 (March 15, 2016): 85–94. http://dx.doi.org/10.21010/ajtcam.v13i3.11.

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Background: Ceratotheca triloba was found to contain three anthraquinones (9, 10-anthracenedione, 1-hydroxy-4-methylanthraquinone and 5, 8-dimethoxy-2, 3, 10, 10a-tetrahydro-1H, 4aH-phenanthrene-4, 9-dione [DTP]) in its roots. Inhibition of the human topoisomerase II enzyme is the basis of some currently used cancer drugs such as doxorubicin which is shown to be cardio-toxic. For this reason we decided to investigate anthraquinones from C. triloba as a possible anticancer drug, however the main limitation was the large quantities of roots that are required to obtain a good yield of the active compound. Therefore the aim of this research was to obtain a higher yield of anthraquinones in hairy roots cultures than the parent plant as well as to compare yields of hairy root, cell suspension and shoot cultures. Materials and Methods: Protocols for seed sterilization, seed germination, shoot cultivation, callus induction, A. rhizogenes mediatedtransformation and hormone supplementations of hairy roots were developed. Results:The results revealed that stem explants was susceptible to transformation by Agrobacterium rhizogenes at a low optical density of 0.2. Induced hairy roots were decontaminated by exposure to cefotaxime at 500mg.l-1 for five days and then 200mg.l-1 for eight days. Visualization of culture extract profiles by TLC revealed anthraquinones were present in all cultures. Analysis of the culture extracts by HPLC showed the highest yield of anthraquinones was produced in hairy root cultures supplemented with 1-Naphthaleneacetic acid [NAA] (8 mg). This was a 17 fold increase compared to field roots (0.47 mg). Conclusion:Therefore C. triloba hairy root cultures are the preferable biological system for anthraquinones production over shoot (0.13 mg) and cell suspension cultures (0.70 mg).

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29

Xu, Zi-Qin, Jing-Fen Jia, and Zhi-De Hu. "Enhancement by Osmotic Treatment of Hairy Root Transformation of Alfalfa Suspension Cultures, and Chromosomal Variation in the Transformed Tissues." Functional Plant Biology 24, no. 3 (1997): 345. http://dx.doi.org/10.1071/pp96032.

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Seeds of alfalfa were germinated on Murashige and Skoog medium without phytohormones. The hypocotyls ofseedlings were excised and cultured on the same medium with 2,4- dichlorophenoxyacetic acid (9.05 mM) to induce callus. Granular calluses were suspended and cultured in Schenk and Hildebrandt medium supplemented with 2.26 mM 2,4-dichlorophenoxyacetic acid on a shaker at 80 rpm. Alfalfa suspension cultures were treated with 0.45 M mannitol for 1 h, and washed twice with 0.16 M CaCl2 .2H2 O. After pretreatment, the pellets were resuspended in Schenk and Hildebrandt medium without phytohormone (10 mL g-1 suspension cultures), and 0.2 mL of Agrobacterium rhizogenes suspension was added. A mixture of alfalfa cells and Agrobacterium was co-cultured at 25 ± 2˚C for 2 days. Transformants (transformed calluses and hairy roots) were obtained by hormone-free selection. Several factors, such as culturing stage of suspension cultures, phytohormone constitution of suspension medium and basal selection medium of transformants, affected the transformation frequency remarkably. Paper electrophoresis revealed that over 70% of transformants could synthesise agropine and mannopine. A comparative cytological analysis revealed the number and structural alterations of chromosomes in the resulting materials. The transformed tissue was not able to be regenerated, possibly due to chromosomal abnormalities.

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30

Kareem, Zana Jamal, Ling Su, Anna Rathgeb, Anke Sirrenberg, Franz Hadacek, Ahmad Hama Ameen H. Rashid, and Petr Karlovsky. "Small-Scale Bioreactor for Sterile Hydroponics and Hairy Roots: Metabolic Diversity and Salicylic Acid Exudation by Hairy Roots of Hyoscyamus niger." Applied Sciences 9, no. 15 (July 28, 2019): 3044. http://dx.doi.org/10.3390/app9153044.

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The volume and complexity of commercial bioreactors for sterile hydroponics and hairy roots are too large for comparative analysis of many cultures. Here a small-scale bioreactor fabricated from standard glass materials and suitable for both airlift and bubble aeration mode is described. The performance of the bioreactor was tested by growing oilseed rape (Brassica napus L.) and rose plants (Rosa canina L.) in sterile hydroponics and by cultivating hairy roots of henbane (Hyoscyamus niger L.) and sesame (Hyoscyamus niger L.). Plants grown in hydroponics for up to six weeks did not show chloroses or necroses. Hairy roots grew faster or comparably fast in bioreactors as compared to shaking flasks. Root exudates of roses and exudates of hairy roots of henbane were subjected to targeted and nontargeted analysis by HPLC coupled with optical and mass spectrometric detectors. The diversity and concentration of hairy root exudates were higher in bioreactors than in shaking flasks. The composition of hairy root exudates of three accessions of H. niger did not match the genetic relatedness among the accessions. Hairy roots of Hyoscyamus niger exuded salicylic acid in amounts varying among plant accessions and between bioreactors and shaking flask cultures.

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31

Эрст (Erst), Анна (Anna) Алексеевна (Аlekseevna), Лариса (Larisa) Николаевна (Nikolayevna) Зибарева (Zibareva), Елена (Elena) Сергеевна (Sergeyevna) Филоненко (Filonenko), and Татьяна (Tat'yana) Витальевна (Vital'yevna) Железниченко (Zheleznichenko). "INFLUENCE METHYL JASMONATE ON PRODUCTION OF ECDYSTEROIDS FROM HAIRY ROOTS OF SILENE LINICOLA C.C.GMELIN." chemistry of plant raw material, no. 4 (December 11, 2018): 159–67. http://dx.doi.org/10.14258/jcprm.2018043807.

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Hairy root cultures are being considered as promising system for producing valuable second metabolites. These genetically transformed root cultures are characterized by high growth rate, genetic stability and growth in hormone free media. Using A. rhizogenes-mediated transformation method (strain А-4), we have obtained hairy root cultures of the ecdysteroid-containing species of Silene linicola. HPLC analysis of the sample studied revealed that 20-hydroxyecdysone (20E), turkesterone and polypodin B were synthesized under the specified experimental conditions. Application of methyl jasmonate at a concentration of 100 μM resulted in stimulation of 20E biosynthesis (up 74%) after 3 days of cultivation and turkesterone up to 35% at 6 days. It was noted that total ecdysteroid content in sample tested varied: turkesterone from 25 to 60%, and 20E from 8 to 30%. At the same time the level of 20E biosynthesis decreased from 0.023 to 0.014% in the samples without methyl jasmonate treatment. Hairy root lines of S. linicola with different responses to the presence of elicitors in the culture medium can be used to study the pathways of ecdysteroid biosynthesis.

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32

Łuczkiewicz, Maria, and Adam Kokotkiewicz. "Genista tinctoria Hairy Root Cultures for Selective Production of Isoliquiritigenin." Zeitschrift für Naturforschung C 60, no. 11-12 (December 1, 2005): 867–75. http://dx.doi.org/10.1515/znc-2005-11-1209.

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Hairy root cultures were established after inoculation of Genista tinctoria in vitro shoots with Agrobacterium rhizogenes, strain ATCC 15834. In transformed roots of G. tinctoria grown in Schenk-Hildebrandt medium without growth regulators the biosynthesis of isoflavones, derivatives of genistein and daidzein, and flavones, derivatives of luteolin and apigenin, characteristic for the intact plant, was completely inhibited. The only compound synthesized in G. tinctoria hairy roots was isoliquiritigenin (2.3 g/100 g DW), a daidzein precursor absent in the intact plant. This compound was stored entirely within cells and it was not until abscisic acid was added (37.8 μm supplement on day 42) that approx. 80% of it was released into the experimental medium. The paper discusses the effect of abscisic acid on the growth of G. tinctoria hairy root cultures, the biosynthesis of isoliquiritigenin and the way it is stored. A prototype basket-bubble bioreactor was designed and built to upgrade the scale of the G. tinctoria hairy root cultures. With immobilized roots and a new aeration system, large amounts of biomass were obtained (FWmax 914.5 g l-1) which produced high contents of isoliquiritigenin (2.9 g/100 g DW). The abscisic acid-induced release of the metabolite from the tissue into the growth medium greatly facilitated subsequent extraction and purification of isoliquiritigenin.

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Berlin, J., I. N. Kuzovkina, C. Rügenhagen, L. Fecker, U. Commandeur, and V. Wray. "Hairy Root Cultures of Peganum harmala II. Characterization of Cell Lines and Effect of Culture Conditions on the Accumulation of ß-Carboline Alkaloids and Serotonin." Zeitschrift für Naturforschung C 47, no. 3-4 (April 1, 1992): 222–30. http://dx.doi.org/10.1515/znc-1992-3-410.

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Abstract Hairy root cultures of Peganum harmala established by genetic transformation with Agrobacterium rhizogenes wild type strains A4, 15834 or carrying the binary vector pLTCgus 1, respectively, were analyzed with respect to their tryptophan-derived metabolites. Beside the previously detected β-carboline alkaloids harmine, harmol, harmaline and harmalol all hairy root cultures were found to contain the β-carboline glucoside ruine and serotonin. The appearance of the root cultures and the levels of β-carboline alkaloids were greatly affected by the nitrogen source of the medium. In the presence of higher levels of ammonium ions (as in MS medium) the roots were short and thick, while in B50 medium (low ammonium) they were thin and long. Ammonium ions had a negative effect on the levels of β-carbolines. The best root cultures accumulated on B50 1.5-2% β-carbolines on a dry mass basis and in the presence of 6% sucrose ca. 10 mg β-carbolines/L x d were formed. In the presence of phytohorm ones root cultures were converted into low producing suspension cultures, from which the highly productive roots were reinduced on phytohormone-free medium.

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Fang, Lingling, Tianhong Yang, and Fabricio Medina-Bolivar. "Production of Prenylated Stilbenoids in Hairy Root Cultures of Peanut (Arachis hypogaea) and Its Wild Relatives A. ipaensis and A. duranensis via an Optimized Elicitation Procedure." Molecules 25, no. 3 (January 24, 2020): 509. http://dx.doi.org/10.3390/molecules25030509.

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Prenylated stilbenoids are phenolic compounds produced in a small number of plants such as peanut (Arachis hypogaea) to counteract biotic and abiotic stresses. In addition to their role in plant defense, they exhibit biological activities with potential application in human health. Whereas non-prenylated stilbenoids such as resveratrol are commercially available, the availability of prenylated stilbenoids is limited. To this end, hairy root cultures of peanut were developed as an elicitor-controlled bioproduction platform for prenylated stilbenoids. An orthogonal array design approach led to the elucidation of an optimized elicitation procedure consisting of co-treatment of the hairy root cultures with 18 g/L methyl-β-cyclodextrin, 125 µM methyl jasmonate, 3 mM hydrogen peroxide (H2O2) and medium supplementation with additional 1 mM magnesium chloride. After 168-h of elicitor treatment, the combined yield of the prenylated stilbenoids arachidin-1, arachidin-2, arachidin-3 and arachidin-5 reached approximately 750 mg/L (equivalent to 107 mg/g DW). Moreover, hairy root cultures from the wild Arachis species A. duranensis and A. ipaensis were developed and shown to produce prenylated stilbenoids upon elicitor treatment. These wild Arachis hairy root lines may provide a platform to elucidate the biosynthetic origin of prenylated stilbenoids in peanut.

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35

Gajurel, Gaurav, Luis Nopo-Olazabal, Emily Hendrix, and Fabricio Medina-Bolivar. "Production and Secretion of Cajaninstilbene Acid in Hairy Root Cultures of Pigeon Pea (Cajanus cajan) Co-Treated with Multiple Elicitors." Plants 11, no. 6 (March 21, 2022): 834. http://dx.doi.org/10.3390/plants11060834.

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Cajaninstilbene acid (CSA) is a prenylated stilbenoid derivative that exhibits neuroprotective, antibacterial, and anti-inflammatory properties. To establish a bioproduction system for this bioactive compound, hairy root cultures of pigeon pea (Cajanus cajan (L.) Millsp.) were developed via Agrobacterium rhizogenes-mediated transformation. The cultures were co-treated with methyl jasmonate, cyclodextrin, hydrogen peroxide, and magnesium chloride to enhance the production of CSA. The amount of CSA that accumulated in the culture medium upon elicitation varied with the period of elicitation. CSA was purified from extracts of the culture medium by semi-preparative HPLC, and its identity was confirmed by tandem mass spectrometry. After 144 h of elicitation in 12-day-old hairy root culture, the total yield of CSA was 8058.618 ± 445.78 μg/g DW, of which approximately 96% was found in the culture medium. The yield of CSA in the elicited hairy root culture was approximately 277-fold higher than in the non-elicited root culture. The difference between the phenotypes of the elicited and non-elicited pigeon pea hairy roots was studied using scanning electron microscopy. The non-elicited hairy roots had uniform surfaces whereas the elicited roots had non-uniform shapes. Pigeon pea hairy roots provide a sustainable platform for producing and studying the biosynthesis of CSA.

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36

Khlebova, L. P., E. S. Brovko, O. V. Bychkova, and N. V. Pavlova. "Optimization of conditions for the induction of Tagetes patula L. hairy roots." Ukrainian Journal of Ecology 9, no. 3 (October 25, 2019): 415–20. http://dx.doi.org/10.15421/2019_119.

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The hairy root cultures are promising sources of secondary metabolites of plants, including rare and endangered species. They possess genetic and biochemical stability, unlimited growth rate in free-hormone medium, short doubling times, high biosynthetic activity and ecological purity of plant raw materials. The hairy root cultures of Tagetes patula L. can be used to produce biologically active substances with biocidal activity. The study aimed to determine the virulent strain of Agrobacterium rhizogenes and the most effective period of co-cultivation of T. patula leaf explants with an agrobacterium to induce actively growing hairy root cultures. We used 3 strains (A-4b, 8196RT and 15834). The time of infection ranged from 3 to 33 hours in increments of 3 hours. We found that 24 h is the best time of infection to induce hairy roots with the highest transformation efficiency (92%). The wild strain A. rhizogenes 15834 turned out to be the most virulent when infected leaf explants of spreading marigold. This strain provided the maximum transformation effect, reaching 85.4%. We have identified 5 actively growing clones of hairy roots with intensive branching, the growth indices of which were 64-75. In the future, they will be transferred to a liquid medium for biomass accumulation and scaling.

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37

Bertóti, Böszörményi, Alberti, Béni, M-Hamvas, Szőke, Vasas, and Gonda. "Variability of Bioactive Glucosinolates, Isothiocyanates and Enzyme Patterns in Horseradish Hairy Root Cultures Initiated from Different Organs." Molecules 24, no. 15 (August 2, 2019): 2828. http://dx.doi.org/10.3390/molecules24152828.

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Horseradish hairy root cultures are suitable plant tissue organs to study the glucosinolate–myrosinase–isothiocyanate system and also to produce the biologically active isothiocyanates and horseradish peroxidase, widely used in molecular biology. Fifty hairy root clones were isolated after Agrobacterium rhizogenes infection of surface sterilized Armoracia rusticana petioles and leaf blades, from which 21 were viable after antibiotic treatment. Biomass properties (e.g. dry weight %, daily growth index), glucosinolate content (analyzed by liquid chromatography-electronspray ionization-mass spectrometry (LC-ESI-MS/MS)), isothiocyanate and nitrile content (analyzed by gas chromatography-mass spectrometry (GC-MS)), myrosinase (on-gel detection) and horseradish peroxidase enzyme patterns (on-gel detection and spectrophotometry), and morphological features were examined with multi-variable statistical analysis. In addition to the several positive and negative correlations, the most outstanding phenomenon was many parameters of the hairy root clones showed dependence on the organ of origin. Among others, the daily growth index, sinigrin, glucobrassicin, 3-phenylpropionitrile, indole-3-acetonitrile and horseradish peroxidase values showed significantly higher levels in horseradish hairy root cultures initiated from leaf blades.

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38

Éva Szőke and Éva Lemberkovics. "Comparative investigation of sesquiterpene components of essential oils originating from intact plants and hairy root chamomile cultures." GSC Advanced Research and Reviews 6, no. 2 (February 28, 2021): 028–49. http://dx.doi.org/10.30574/gscarr.2021.6.2.0016.

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The importance of chamomile (Chamomilla recutita) inflorescence is widely known in classical and folk medicine, with the largest group of its effective constituents forming the essential oil (chamazulene, a-bisabolol, α-farnesene, trans-β-farnesene, spathulenol, cis/trans-en-in-dicycloethers). Among cultivated species, the Hungarian BK-2 contains more chamazulene in its essential oil than the German Degumil type, which is mainly cultivated for its a-bisabolol. Both components have important antiinflammatory activities. Wild populations can be easily distinguished from cultivated ones by their high amount of bisaboloides, particularly the flower of Hungarian Szabadkígyós wild type, which contained on average 48 % of the biologically active (-)-a-bisabolol. The population of Szabadkígyós has good salt tolerance which is important owing to global warming, because the proportion of saline areas is increasing worldwide. To keep the genome of Szabadkígyós having high (-)-a-bisabolol content, Szőke and research team used biotechnological methods. Sterile plantlets, were infected by Agrobacterium rhizogenes strains #A-4, #15834, #R-1601. The hairy root clones possessing the best growing and biosynthetical potential were multiplied for phytochemical investigations. Pharmacologically important compounds of their essential oils were followed in great detail. The amount of in vitro cultured terpenoids and polyin compounds was compared with that of in vivo plants. GC-MS studies showed that sterile chamomile cultures generated the most important terpenoid and polyin compounds characteristics of the mother plant. Berkheyaradulene, geranyl-isovalerat and cedrol as new components were identified in these sterile cultures. The main component of hairy root cultures (D/400, D/1, D/100 and Sz/400) was tr-b-farnesene and in addition one new compound: a-selinene was identified. Hairy root culture originated from chamomile collected in Szabadkígyós was intensive increased the essential oil content and pharmacological active compounds: (-) -α-bisabolol and β-eudesmol was also synthetized in large quantity. Furthermore, in vitro organized cultures were made from this population to obtain propagation material containing numerous active substances.

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Vinterhalter, Branka, Vladimir Orbović, and Dragan Vinterhalter. "Transgenic root cultures of Gentiana punctata L." Acta Societatis Botanicorum Poloniae 68, no. 4 (2014): 275–80. http://dx.doi.org/10.5586/asbp.1999.038.

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Shoot cultures of Gentiana punctata L. were inoculated with suspension of Agrobacterium rhizogenes strain A4 M70GUS. Hairy roots which appeared 2-3 weeks later were cultured on hormone-free, liquid, WPM (Lloyd and McCown 1980) basal medium for more than 5 years (60 subcultures). Growth rate of transformed roots was higher than the growth rate of nontransformed roots. Spontaneous shoot regeneration occured only in three culture vessels in subcultures No. 40 and 42. Plants had phenotype characteristics typical for A. rhizogenes transformed plants including: wrincled leaves, short internodes, plagiotropic roots and in general their growth rate was reduced. These plants also manifested precocious formation of flower buds without vernalization and flowering under in vitro conditions. Flowers were pale yellow, the same as in the standard phenotype.

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40

Akhgari, Amir, Into Laakso, Hannu Maaheimo, Young Hae Choi, Tuulikki Seppänen-Laakso, Kirsi-Marja Oksman-Caldentey, and Heiko Rischer. "Methyljasmonate Elicitation Increases Terpenoid Indole Alkaloid Accumulation in Rhazya stricta Hairy Root Cultures." Plants 8, no. 12 (November 22, 2019): 534. http://dx.doi.org/10.3390/plants8120534.

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Methyl jasmonate is capable of initiating or improving the biosynthesis of secondary metabolites in plants and therefore has opened up a concept for the biosynthesis of valuable constituents. In this study, the effect of different doses of methyl jasmonate (MeJA) elicitation on the accumulation of terpenoid indole alkaloids (TIAs) in the hairy root cultures of the medicinal plant, Rhazya stricta throughout a time course (one-seven days) was investigated. Gas chromatography-mass spectrometry (GC-MS) analyses were carried out for targeted ten major non-polar alkaloids. Furthermore, overall alterations in metabolite contents in elicited and control cultures were investigated applying proton nuclear magnetic resonance (1H NMR) spectroscopy. Methyl jasmonate caused dosage- and time course-dependent significant rise in the accumulation of TIAs as determined by GC-MS. The contents of seven alkaloids including eburenine, quebrachamine, fluorocarpamine, pleiocarpamine, tubotaiwine, tetrahydroalstonine, and ajmalicine increased compared to non-elicited cultures. However, MeJA-elicitation did not induce the accumulation of vincanine, yohimbine (isomer II), and vallesiachotamine. Furthermore, principal component analysis (PCA) of 1H NMR metabolic profiles revealed a discrimination between elicited hairy roots and control cultures with significant increase in total vindoline-type alkaloid content and elevated levels of organic and amino acids. In addition, elicited and control samples had different sugar and fatty acid profiles, suggesting that MeJA also influences the primary metabolism of R. stricta hairy roots. It is evident that methyl jasmonate is applicable for elevating alkaloid accumulation in “hairy root” organ cultures of R. strica.

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41

Barr, D. J. S., and N. L. Désaulniers. "The life cycle of Lagena radicicola, an oomycetous parasite of wheat roots." Canadian Journal of Botany 68, no. 4 (April 1, 1990): 813–24. http://dx.doi.org/10.1139/b90-108.

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Lagena radicicola Vanterpool & Ledingham is an obligate parasite inside root hairs and epidermal cells. It was cultured in a unifungal state on wheat in pots. The life cycle was examined by both light and transmission electron microscopy. The thallus developed inside a single host cell and formed either a single sporangium or one to four resting spores. Zoospore cleavage was completed in vesicles outside the root. The resting spores were similar to oospores in their development and cytology, but there was no evidence of cell fusion and sexuality. Virus-like particles were seen in 3- to 12-month-old cultures, and infected cells became degenerate. Key words: Oomycetes, ultrastructure, virus-like particles, biocontrol.

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42

Roy, Arpita. "Hairy Root Culture an Alternative for Bioactive Compound Production from Medicinal Plants." Current Pharmaceutical Biotechnology 22, no. 1 (December 31, 2020): 136–49. http://dx.doi.org/10.2174/1389201021666201229110625.

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: Medicinal plants produce a diverse group of phytocompounds like anthraquinones, alkaloids, anthocyanins, flavonoids, saponins, and terpenes which are used in pharmaceutical, perfume, cosmetics, dye and flavor industries. Commercial source of these metabolites is field-grown plants, which are generally influenced by seasonal changes. Biotechnology possesses a significant role in production of high-value secondary metabolites. By incorporating biotechnological methods, it is feasible to manage biosynthetic pathways of the plant to enhance phytocompound production that is of pharmaceutical interest. Plant cell suspension, shoot, adventitious root and hairy root culture are considered as alternative methods for important bioactive compound production. These methods are controllable, sustainable and overcome several inconveniences for large scale secondary metabolites production. At present research on hairy root culture for valuable bioactive compound production has gained a lot of attention. Agrobacterium rhizogenes is an agent which causes hairy root disease in a plant and this leads to the neoplastic growth of root which is characterized by higher growth rate and genetic stability. Various studies explore the hairy root culture for production of a wide range of bioactive compounds. Scale-up of hairy root culture using bioreactors has provided an opportunity to enhance bioactive compound production at the commercial level. The present review discusses the role of hairy root culture in the production of valuable bioactive compounds, the effect of culture parameters on bioactive compound production and bioreactor applications.

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Zayed, Rawia, and Michael Wink. "Induction of Pyridine Alkaloid Formation in Transformed Root Cultures of Nicotiana tabacum." Zeitschrift für Naturforschung C 64, no. 11-12 (December 1, 2009): 869–74. http://dx.doi.org/10.1515/znc-2009-11-1219.

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Hairy root cultures of Nicotiana tabacum were set up by excised root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analyzing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of pyridine alkaloids, mainly nicotine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid, in order to stimulate the biosynthesis of pyridine alkaloids. Profile and amounts of pyridine alkaloids were analyzed using capillary GLC-MS. Treatment of the cultures with methyljasmonate (50 μM) increased the alkaloid accumulation ca. 7-fold up to a level of 0.58 mg/g fresh weight as compared to untreated controls. Quercetin (200 μM) enhanced the alkaloid production ca. 4-fold (0.34 mg/g fresh weight) within 24 h. In contrast salicylic acid in all tested concentrations decreased the alkaloid level to 1 μg/g fresh weight. Also the inhibitory effect of salicylic acid on the elicitation effect of methyljasmonate and quercetin was investigated.

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Zayed, Rawia, and Michael Wink. "Induction of Tropane Alkaloid Formation in Transformed Root Cultures of Brugmansia suaveolens (Solanaceae)." Zeitschrift für Naturforschung C 59, no. 11-12 (December 1, 2004): 863–67. http://dx.doi.org/10.1515/znc-2004-11-1216.

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Hairy root cultures of Brugmansia suaveolens were set up by infection of root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analysing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of tropane alkaloids, such as hyoscyamine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid in order to stimulate the biosynthesis of tropane alkaloids. Profile and amounts of tropane alkaloids were analysed using capillary GLC-MS. At least 18 different tropane alkaloids could be identified. Treatment of the cultures with 200 μᴍ methyljasmonate increased the alkaloid accumulation 25-fold up to a level of 1 mg/g fresh weight as compared to untreated controls. Quercetin enhanced the alkaloid production 10 fold (0.4 mg/g fresh weight) within 24 h. In contrast 100 μᴍ salicylic acid decreased alkaloids to a level of 1 μg/g fresh weight.

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Vergara-Martínez, Victor M., Samuel E. Estrada-Soto, Susana Valencia-Díaz, Karlina Garcia-Sosa, Luis Manuel Peña-Rodríguez, José de Jesús Arellano-García, and Irene Perea-Arango. "Methyl jasmonate enhances ursolic, oleanolic and rosmarinic acid production and sucrose induced biomass accumulation, in hairy roots of Lepechinia caulescens." PeerJ 9 (April 27, 2021): e11279. http://dx.doi.org/10.7717/peerj.11279.

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Background Ursolic (UA), oleanolic (OA) and rosmarinic (RA) acids are bioactive metabolites found in Lepechinia caulescens that have generated interest for their health benefits, which include antimicrobial, antioxidant, antimutagenic, gastroprotective, antidiabetic, antihypertensive and anti-inflammatory properties, among others. To date, very few attempts have been made to evaluate the potential for simultaneous production of these bioactive compounds, using a biotechnological approach. Hairy root cultures offer a biotechnology approach that can be used to study the factors affecting the biosynthesis and the production of UA, OA and RA. In the current study, we established hairy root cultures of L. caulescens and evaluated the effect of sucrose on biomass accumulation, and the effect of different concentrations and times of exposure of methyl jasmonate (MeJA), on the accumulation of UA, OA and RA. Methods Leaves from plants of L. caulescens were inoculated with Agrobacterium rhizogenes strain ATCC 15834. PCR of rolB gene confirmed the transgenic nature of hairy roots. Hairy roots were subcultured in semisolid MSB5 medium, supplemented with 15, 30, 45 or 60 g/L sucrose and after 4 weeks, dry weight was determined. The accumulation of UA, OA and RA of wild plants and hairy roots were determined by HPLC. Finally, the hairy roots were treated with 0, 100, 200 and 300 µM of MeJA and the content of bioactive compounds was analyzed, after 24, 48 and 72 h. Results High frequency transformation (75%) was achieved, using leaf explants from axenic seedlings, infected with A. rhizogenes. The hairy roots showed an enhanced linear biomass accumulation, in response to the increase in sucrose concentration. The hairy root cultures in MSB5 medium, supplemented with 45 g/L sucrose, were capable to synthesizing UA (0.29 ± 0.00 mg/g DW), OA (0.57 ± 0.00 mg/g DW) and RA (41.66 ± 0.31 mg/g DW), about two, seven and three times more, respectively, than in roots from wild plants. Elicitation time and concentration of MeJA resulted in significant enhancement in the production of UA, OA and RA, with treatments elicited for 24 h, with a concentration of 300 µM of MeJA, exhibiting greatest accumulation. Conclusion This is the first report on development of hairy root cultures of L. caulescens. Future studies should aim towards further improving triterpenes and polyphenolic compound production in hairy roots of L. caulescens, for use in the pharmaceutical and biotechnological industry.

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Berlin, J., C. Rügenhagen, M. Rippert, and S. Erdogan. "Effects of Culture Conditions on Isoflavonoid Levels of Transformed and Non-Transformed Cultures of Lupinus -a Comparison of Suspension and Hairy Root Cultures." Zeitschrift für Naturforschung C 46, no. 9-10 (October 1, 1991): 735–42. http://dx.doi.org/10.1515/znc-1991-9-1004.

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Abstract Some highly productive suspension and hairy root cultures were found among several transformed cultures of Lupinus polyphyllus and L. hartwegii. A transformed suspension culture Lupo 30150 and a root culture Luha 15834 containing the highest specific isoflavone glucoside content were characterized and compared with normal phytohormone-dependent lines with respect to product stability as well as to their responsiveness to external triggers, e.g. response to changes in the medium. While phytohormone-dependent suspension cultures lost their initial ability to form increased levels of isoflavonoids on phytohormone-free medium, the transformed phytohorm one-independent suspension Lupo 30150 remained a highly productive line, despite the fact that its specific levels decreased to 60% of the initial values during several years in liquid medium. Highest stability of product patterns and levels were noted for the transformed root culture. Phytohorm ones had little effect on growth and isoflavonoid levels in suspension cultures, while they reduced both strongly in root cultures. In the presence of 2,4-D the root culture changed into an aggregated low producing suspension culture from which the root state was recovered on phytohorm one-free medium. As long as the root state was main tained, isoflavonoid levels could not be distinctly improved by media variation while specific isoflavonoid levels of suspensions were increased by stress factors such as phosphate depletion. When suspensions were transferred to fresh medium phenylalanine ammonia-lyase was greatly induced within 24 h, while the activity remained nearly unchanged in root cultures.

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Yeon Kwon, Do, Haeng Hoon Kim, Jong Seok Park, Sang Un Park, and Nam Il Park. "Production of Bacalin, Bacalein and Wogonin in Hairy Root Culture of American Skullcap (Scutellaria lateriflora)by Auxin Treatment." Biosciences, Biotechnology Research Asia 14, no. 2 (June 25, 2017): 673–77. http://dx.doi.org/10.13005/bbra/2493.

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ABSTRACT: The hairy root culture of American Skullcap (Scutellaria lateriflora) was studied to investigate the biomass and flavonoids content (baicalin, baicalein and wogonin) in response of various auxin concentrations.The growth rates of the hairy roots varied significantly only at IBA 0.1 mg/L and for all other auxin treatments did not vary significantly. The biomass of hairy roots was 8% higher when treated with IBA 0.1 mg/L and biomass was almost similar and slightly lower levels when treated with various IAA concentration and NAA, respectively. However, the auxins treatments responsed positively to increase flavone production in American Skullcaphairy root culture. The auxin indole-3-butyric acid (IBA) at 1 mg/L performed the best for the accumulation of baicalin and wogonin. The auxin IBA at 1 mg/L accumulated 1.64 and 2.92 times higher baicalin and wogonin, respectively compared to control treatment. Meanwhile, the highest levels of baicalein were observed for hair root cultures in the presence of 1-naphthaleneacetic acid (NAA) at 0.1 mg/L achieving 2.38 times higher than that of accumulated in the control. These findings indicate that hairy root cultures of S. lateriflorausing liquid 1/2MS medium supplemented with auxin could be a valuable alternative approach for flavonoid production.

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Park, Sang, Xiaohua Li, Seok Eom, Chung Lee, and Sook Lee. "E-p-methoxycinnamic acid production in hairy root cultures of Scrophularia buergeriana Miquel." Archives of Biological Sciences 62, no. 3 (2010): 649–52. http://dx.doi.org/10.2298/abs1003649p.

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E-p-methoxycinnamic acid (MCA) is one of the main active constituents of Scrophularia buergeriana Miquel and has hepatoprotective, anti-amnestic, and neuroprotective activities. For studying in vitro production of MCA, we established a hairy root culture of S. buergeriana by infecting leaf explants with Agrobacterium rhizogenes R1000, and tested the growth and MCA production of these cultures using different strengths of media and concentrations of auxins. Hairy roots grown in half-strength MS medium showed the highest levels of growth (10.3 g/l) as well as MCA production (0.83 mg/100 g dry weight). Hairy root culture with the treatment of 0.5 mg/l IBA produced the highest amount of dry weight (11.8 g/l) and MCA (1.26 mg/100 g dry weight) production. These results demonstrate that the hairy root culture of S. buergeriana is a valuable alternative approach for the production of MCA. .

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Torkamani, Mohammad, Morad Jafari, Nasser Abbaspour, Reza Heidary, and Naser Safaie. "Enhanced production of valerenic acid in hairy root culture of Valeriana officinalis by elicitation." Open Life Sciences 9, no. 9 (September 1, 2014): 853–63. http://dx.doi.org/10.2478/s11535-014-0320-3.

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AbstractValerenic acid (VA) is a pharmacologically-active sesquiterpene found in valerian (Valeriana officinalis L., Valerianaceae) roots and rhizomes. The plant produces only small amounts of this metabolite naturally. So, induction of hairy roots as well as elicitation can be useful to increase its commercial production. In this study, Wild-type strain ‘A13’ of Agrobacterium rhizogenes was used to induce hairy roots in valerian. The influence of three different elicitors including Fusarium graminearum extract (FE), methyl jasmonate (MJ) and salicylic acid (SA) on VA production in the selected hairy root line ‘LeVa-C4’ was also investigated. The 23-day-old cultures were treated with different concentrations of the elicitors at exposure time of 3 and 7 days. FE (1%) and MJ (100 µM L−1) highly promoted VA production at 7 days after elicitation, to a level of 12.31- and 6-fold higher than that of non-elicited controls, respectively, and FE did not exert any negative effects on biomass yield of hairy root. SA did not significantly increase the production of VA. This is the first time study to assess the elicitation of hairy root cultures to promote VA biosynthesis in valerian and the resulting experiments demonstrated that F. graminearum extract and MJ were indeed a potent inducer of VA biosynthesis.

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Tappe, R., H. Budzikiewicz, I. Ionkova, and A. W. Alfermann. "Triterpene glycosides from transformed root cultures ofAstragalus mongholicusBGE." Spectroscopy 12, no. 1 (1994): 1–8. http://dx.doi.org/10.1155/1994/308939.

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Transfonned root cultures ofAstragalus mongholicusBge. have been established by infecting of sterile grown plants with AgrobacterilDfl rhizogenes strains LBA 9402 and ATCC 15834. A promoting effect of acetyl syringone on the Agrobacteriwn-mediated hairy root (HR) initiation was obselVed. The transfonned root clones were assayed for opines by electrophoresis and 1LC. All of them were positive for agropine/mannopine. It was shown, that Astragalus-hairy roots (HR) of the transfonned cultures produce the same kind of saponins in considerable amounts as those of intact plants. The triterpene saponins have been isolated from the methanol extract of the HR and identified as cycloartane derived saponins. On the basis of analytical data obtained by mass spectrometry (NI-F AB-MS, EI-MS), and homo- and heteronuclear NMR experiments (DEPT, DQF-COSY, HOHAHA, ROESY, HMQC, HMBC), the structures of three triterpenoid glycosides were determined as astragaloside I, IT, ill [1,2]. A complete assignment of the IH_ and 13C-NMR data is given.

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Journal articles on the topic 'Hairi root cultures'

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