Mitragyna speciosa: Hairy Root Culture for Triterpenoid Production and High Yield of Mitragynine by Regenerated Plants

2008 ◽  
Vol 63 (9-10) ◽  
pp. 691-698 ◽  
Author(s):  
Siriwan Phongprueksapattana ◽  
Waraporn Putalun ◽  
Niwat Keawpradub ◽  
Juraithip Wungsintaweekul
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
High Yield ◽  
Naphthaleneacetic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Mitragyna Speciosa ◽  
In Vitro Plantlets ◽  
Regenerated Plants

Hairy root cultures of Mitragyna speciosa were established by infection of Agrobacterium rhizogenes ATCC 15834 and maintained in McCown woody plant medium (WPM) supplemented with 0.5 mg/l naphthaleneacetic acid. The hairy roots were identified for the rooting genes loci of rolA and rolB by polymerase chain reaction. For studying the secondary metabolite production, the n-hexane extract of the hairy roots was prepared and the compounds were isolated by silica gel column chromatography, affording triterpenoids (ursolic acid and oleanolic acid) and phytosterols (β-sitosterol and stigmasterol). The shoots from the hairy root cultures were regenerated and differentiated to the plantlets. For micropropagation, shoot multiplication was successfully induced from the axillary buds of the regenerated plantlets in WPM supplemented with 0.1 mg/l thidiazuron. The mitragynine contents of 5-monthold regenerated plants and in vitro plantlets (germinated from seeds) were determined using the TLC-densitometric method. The regenerated plants contained (14.25 ± 0.25) mg/g dry wt mitragynine, whereas the in vitro plantlets contained (4.45 ± 0.09) mg/g dry wt.

scholarly journals Alterations in oleanolic acid and sterol content in marigold (Calendula officinalis) hairy root cultures in response to stimulation by selected phytohormones

2021 ◽  
Vol 43 (3) ◽  
Author(s):  
Abdulwadood S. M. Alsoufi ◽  
Klaudia Staśkiewicz ◽  
Michał Markowski
Keyword(s):  
Oleanolic Acid ◽  
Hairy Root ◽  
Hairy Roots ◽  
Naphthaleneacetic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Calendula Officinalis ◽  
Plant Metabolites ◽  
Triterpenoid Saponins

AbstractHairy root cultures are an efficient tool for the biotechnological production of plant metabolites and a convenient experimental model for analyzing the effect of various compounds on plant metabolism. In contrast to many other types of in vitro plant cultures, hairy roots do not require an external supply of phytohormones to the medium. Consequently, plant growth regulators such as auxins and cytokinins are rarely used as elicitors in hairy root in vitro cultures; however, they can strongly influence plant defense responses. The aim of this study was to investigate the influence of two auxins: natural indole-3-acetic acid (IAA) and synthetic 1-naphthaleneacetic acid (NAA), as well as two cytokinins: natural kinetin and synthetic 6-benzylaminopurine (BAP) at a concentration of 0.75 mg/L on the metabolism of sterols and triterpenoids in Calendula officinalis hairy roots. Auxins prevented the accumulation of triterpenoid saponins (oleanolic acid glycosides), while cytokinin BAP increased their accumulation by 17% and their release into the culture medium by a factor of 10. Other cytokinins and kinetins increased the sterol levels by 17%, the level of stigmasterol by 15%, and the level of isofucosterol by 7 times.

scholarly journals Production of Bacalin, Bacalein and Wogonin in Hairy Root Culture of American Skullcap (Scutellaria lateriflora)by Auxin Treatment

2017 ◽  
Vol 14 (2) ◽  
pp. 673-677 ◽  
Author(s):  
Do Yeon Kwon ◽  
Haeng Hoon Kim ◽  
Jong Seok Park ◽  
Sang Un Park ◽  
Nam Il Park
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Control Treatment ◽  
Naphthaleneacetic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Alternative Approach ◽  
Scutellaria Lateriflora

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.

scholarly journals Production of Triterpenoid Sapogenins in Hairy Root Cultures of Silene vulgaris

2015 ◽  
Vol 10 (11) ◽  
pp. 1934578X1501001
Author(s):  
Yeon Bok Kim ◽  
Darwin W. Reed ◽  
Patrick S. Covello
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Leaf Explants ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Silene Vulgaris ◽  
In Vitro Production ◽  
Triterpenoid Saponins ◽  
Meja Treatment

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.

Sustainable bioproduction of phytochemicals by plant in vitro cultures: anticancer agents

2005 ◽  
Vol 3 (2) ◽  
pp. 90-100 ◽  
Author(s):  
Michael Wink ◽  
A. Wilhelm Alfermann ◽  
Rochus Franke ◽  
Bernhard Wetterauer ◽  
Melanie Distl ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Anticancer Agents ◽  
Complex Structure ◽  
Suspension Cultures ◽  
Cell Suspensions ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Commercial Scale

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.

scholarly journals Effect of Auxins on Anthocyanin Accumulation in Hairy Root Cultures of Tartary Buckwheat Cultivar Hokkai T10

2016 ◽  
Vol 11 (9) ◽  
pp. 1934578X1601100 ◽  
Author(s):  
Chang Ha Park ◽  
Aye AyeThwe ◽  
Sun Ju Kim ◽  
Jong Seok Park ◽  
MariadhasValan Arasu ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Tartary Buckwheat ◽  
Naphthaleneacetic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Indolebutyric Acid ◽  
Liquid Chromatograph

This study investigated the influence of auxins on the growth of hairy roots and accumulation of anthocyanins, including cyanidin 3- O-glucoside (C3gl) and cyanidin 3- O-rutinoside (C3r), in the hairy root culture of tartary buckwheat cultivar Hokkai T10. C3gl and C3r contents were evaluated using highperformance liquid chromatograph (HPLC). Four auxins, 2,4-dichlorophenoxyacetic acid (2,4-D), indoleacetic acid (IAA), indolebutyric acid (IBA) and naphthaleneacetic acid (NAA), were added to the medium of the hairy root cultures at diverse concentrations. IAA, IBA and 2,4-D promoted the growth of hairy roots since the dry weight of the roots was slightly higher than or comparable with that of the control. However, NAA at all concentrations suppressed the growth of hairy roots. Generally, auxin treatments resulted in higher accumulation of C3gl and C3r than that of the control except for 2.85 μM IAA and 2.69 μM NAA. The amount of C3gl and C3r after treatment with 4.92 μM IBA was the highest among all treatments and was 3.24 times more than that of the control. Our results suggested that auxins at appropriate concentrations might facilitate hairy root growth of tartary buckwheat and enhance the production of C3gl and C3r.

scholarly journals Preliminary results of indole alkaloids production in different roots of Catharanthus roseus cultured in vitro

2014 ◽  
Vol 70 (4) ◽  
pp. 261-265 ◽  
Author(s):  
Agnieszka Pietrosiuk ◽  
Mirosława Furmanowa
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Indole Alkaloids ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Preliminary Results ◽  
Intact Plants

Six groups of untransformed and hairy root cultures of <em>Catharunthus roseus</em> (L.) G. Don were established. <em>Agrobacterium rhizogenes</em> strains: ATCC 15834, LBA 9403, and TR 105 were used for infection of the 3-week old rooted plantlets of <em>C. roseus</em>. The highest contents of examined indole alkaloids were found in: roots of intact plants - yohimbine and serpentine; in hairy roots - catharanthine. Vinblastine and ajmalicine were detected in untransformed roots of plants regenerated in vitro, and transferred to the soil for 5 months.

Clone selection of in vitro horseradish hairy root cultures

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
R Bertóti ◽  
Á Alberti ◽  
A Böszörményi ◽  
R Könye ◽  
T Horváth ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Clone Selection ◽  
Selection Of

scholarly journals Carrot hairy roots: factories for secondary metabolite production

2020 ◽  
Vol 71 (22) ◽  
pp. 6861-6864
Author(s):  
María A Pedreño ◽  
Lorena Almagro
Keyword(s):  
Phenolic Compounds ◽  
Secondary Metabolite ◽  
Hairy Root ◽  
Hairy Roots ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Black Carrot ◽  
Metabolite Production ◽  
Experimental Botany ◽  
Induced Changes

This article comments on: Barba-Espín G, Chen S-T, Agnolet S, Hegelund JN, Stanstrup J, Christensen JH, Müller R, Lütken H. 2020. Ethephon-induced changes in antioxidants and phenolic compounds in anthocyanin-producing black carrot hairy root cultures. Journal of Experimental Botany 71, 7030–7045.

Hairy Roots

2021 ◽  
pp. 219-252
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Growth ◽  
Root Culture ◽  
Hairy Root Culture ◽  
High Growth Rate ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

scholarly journals Hairy Root Cultures of Eurycoma longifolia and Production of Anti-inflammatory 9-Methoxycanthin-6-one

2018 ◽  
Vol 13 (5) ◽  
pp. 1934578X1801300
Author(s):  
Trang Thu Tran ◽  
Nam Trung Nguyen ◽  
Ngoc Bich Pham ◽  
Huy Nhat Chu ◽  
Trong Dinh Nguyen ◽  
...  
Keyword(s):  
Hairy Root ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Peritoneal Macrophages ◽  
High Yield ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Eurycoma Longifolia ◽  
Anti Inflammatory

The content of bioactive alkaloid 9-methoxycanthin-6-one in the roots of Eurycoma longifolia is rather small. In the current study, a high yield of 9-methoxycanthin-6-one was obtained from transgenic hairy roots (1.139 ± 0.20% dry weight, DW) compared to wild roots (0.164 ± 0.25% DW) of E. longifolia. Three liquid basal media supplemented with 3% sucrose were tested for the growth and accumulation of 9-methoxycanthin-6-one. The fastest growth was obtained in the Shenck and Hildebrandt (SH) medium while the highest 9-methoxycanthin-6-one content was observed in McCown's woody plant (WP) medium. In this study, compared to controls, the production of 9-methoxycanthin-6-one was increased by the maximum of 2.6 and 4.0 times after the addition of jasmonic acid and yeast extract, respectively, into hairy root cultures. In addition, pro-inflammatory cytokines IL-6 and TNF-α were significantly inhibited by 9-methoxycanthin-6-one in lipopolysaccharide-stimulated murine RAW264.7 cells, peritoneal macrophages and human THP-1 macrophages These results suggest that the elicited hairy root culture of E. longifolia is an alternative system for the production of an abundant source of anti-inflammatory 9-methoxycanthin-6-one.

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