scholarly journals Production of oleanolic acid glycosides by hairy root established cultures of Calendula officinalis L.

2013 ◽  
Vol 60 (3) ◽  
Author(s):  
Marek Długosz ◽  
Ewa Wiktorowska ◽  
Anita Wiśniewska ◽  
Cezary Pączkowski
Keyword(s):  
Oleanolic Acid ◽  
Hairy Root ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Calendula Officinalis ◽  
Kinase Gene ◽  
High Positive Correlation ◽  
Oleanolic Acid Glycosides ◽  
Calendula Officinalis L

In order to initiate hairy root culture initiation cotyledons and hypocotyls of Calendula officinalis L. were infected with Agrobacterium rhizogenes strain ATCC 15834 or the same strain containing pCAMBIA 1381Z vector with β-glucuronidase reporter gene under control of promoter of NIK (Nematode Induced Kinase) gene. The efficiency of induction of hairy roots reached 33.8% for cotyledons and 66.6% for hypocotyls together for both transformation experiments. Finally, eight control and nine modified lines were established as a long-term culture. The hairy root cultures showed the ability to synthesize oleanolic acid mainly (97%) as glycosides; control lines contained it at the average 8.42 mg · g(-1) dry weight in tissue and 0.23 mg · dm(-3) in medium; modified lines: 4.59 mg · g(-1) for the tissue, and 0.48 mg · dm(-3) for the medium. Additionally lines showed high positive correlation between dry/fresh weight and oleanolic acid concentration in tissue. Using the Killiani mixture in acidic hydrolysis of oleanolic acid glycosides released free aglycones that were partially acetylated in such conditions.

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.

Polyprenols in hairy roots of Coluria geoides

2000 ◽  
Vol 28 (6) ◽  
pp. 790-791 ◽  
Author(s):  
K. Skorupińska-Tudek ◽  
V. S. Hung ◽  
O. Olszowska ◽  
M. Furmanowa ◽  
T. Chojnacki ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Lipid Content ◽  
Mammalian Cells ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Living Organisms ◽  
Long Chain

Long-chain polyisoprenoid alcohols built from several up to more than 100 isoprenoid units are common constituents of all living organisms. They were found mostly in plants, bacteria, yeasts and mammalian cells. In vitro hairy root culture of Coluria geoides was obtained from plants transformed with Agrobacterium rhizogenes. Growth was optimal at 0.75% (w/v) glucose and at 22 °C. Dry samples of roots were extracted and lipid content was analysed by HPLC. According to our estimation, polyprenols are accumulated in roots of C. geoides cultivated in vitro as a mixture of several prenologues with the dominating prenol composed of 16 isoprenoid units. The content of polyprenols in tissue was approx. 300 μg/g of dry weight.

Biosynthesis of oleanolic acid glycosides in protoplasts isolated from Calendula officinalis L. roots

2006 ◽  
Vol 28 (3) ◽  
pp. 217-223 ◽  
Author(s):  
Dariusz Ruszkowski ◽  
Anna Szakiel ◽  
Elbieta Auguciska ◽  
Wirginia Janiszowska
Keyword(s):  
Oleanolic Acid ◽  
Calendula Officinalis ◽  
Oleanolic Acid Glycosides ◽  
Calendula Officinalis L

Devil’s Claw Hairy Root Culture in Flasks and in a 3-L Bioreactor: Bioactive Metabolite Accumulation and Flow Cytometry

2010 ◽  
Vol 65 (7-8) ◽  
pp. 472-478 ◽  
Author(s):  
Vediha Homova ◽  
Jost Weber ◽  
Josef Schulze ◽  
Kalina Alipieva ◽  
Thomas Bley ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Hairy Root ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Water Soluble ◽  
Root Cultures ◽  
Phenylethanoid Glycosides ◽  
Devil’S Claw ◽  
Shake Flasks

Phenylethanoids are a group of natural water-soluble compounds with high biological value, which could potentially be commercially produced by hairy root cultures. Thus, we have examined the capacity of transformed root cultures of Devil’s claw (Harpagophytum procumbens) to accumulate four phenylethanoid glycosides - β-OH-verbascoside, verbascoside, leucosceptoside A, and martynoside - in shake-flasks and a 3-L stirred tank reactor. Verbascoside was found to be the major phenylethanoid, and its maximal contents were the same (1.12 mg/g dry weight) in both kinds of culture. However, peak leucosceptoside A contents were 1.6-times higher in bioreactor cultures than in shake-flask cultures. Flow cytometry analysis revealed that G0 + G1-phase cells predominated throughout the growth of the cultures, which was in accordance with the very high proportion of quiescent cells in the transformed roots. The results provide the first demonstration of the potential utility of Devil’s claw hairy roots as biofactories for producing high-value phenylethanoid glycosides.

Diterpenoid Production in Hairy Root Culture of Salvia sclarea L.

2008 ◽  
Vol 63 (7-8) ◽  
pp. 621-624 ◽  
Author(s):  
Łukasz Kuzma ◽  
Elzbieta Bruchajze ◽  
Halina Wysokińska
Keyword(s):  
Liquid Medium ◽  
Hairy Root ◽  
Total Content ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Growth Cycle ◽  
Transformed Roots ◽  
Salvia Sclarea ◽  
Half Strength

Growth and diterpenoid accumulation (salvipisone, ferruginol, aethiopinone and 1-oxoaethiopinone) during the growth cycle of a Salvia sclarea hairy root culture are described. The roots transformed by Agrobacterium rhizogenes (LBA 9402) were cultured in half-strength B5 liquid medium supplemented with 30 g L-1 sucrose under light (16 h/8 h light/dark). A culture period of 30 days was optimal for both biomass and diterpenoid production. The total content of four diterpenoids in the hairy roots [(27.3 ± 0.6) mg g-1 dry weight] was higher than that of roots of field-grown S. sclarea plants [(3.15 ± 0.15) mg g-1 dry weight]. In transformed roots, aethiopinone was the main diterpenoid, whereas the principal diterpenoid of natural roots was salvipisone.

scholarly journals E-p-methoxycinnamic acid production in hairy root cultures of Scrophularia buergeriana Miquel

2010 ◽  
Vol 62 (3) ◽  
pp. 649-652 ◽  
Author(s):  
Sang Park ◽  
Xiaohua Li ◽  
Seok Eom ◽  
Chung Lee ◽  
Sook Lee
Keyword(s):  
Hairy Root ◽  
Leaf Explants ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
In Vitro Production ◽  
Methoxycinnamic Acid ◽  
Half Strength ◽  
Vitro Production

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. .

scholarly journals Response Surface Modelling of Noradrenaline Production in Hairy Root Culture of Purslane (Portulaca oleracea L.)

2015 ◽  
Vol 3 (6) ◽  
pp. 349 ◽  
Author(s):  
Mehdi Ghorbani ◽  
Aghil Ghorbani ◽  
Mansoor Omidi ◽  
Seyed Mohammad Hashemi
Keyword(s):  
Response Surface ◽  
Hairy Root ◽  
Root Culture ◽  
Dry Weight ◽  
Hairy Root Culture ◽  
Portulaca Oleracea ◽  
Noradrenaline Content ◽  
Hairy Root Line ◽  
Response Surface Modelling ◽  
Common Purslane

Common purslane (Portulaca oleracea L.) is an annual plant as one of the natural sources for noradrenaline hormone. In this research, hairy root culture of purslane was established by using Agrobacterium rhizogenes strain ATCC 15834. In the following, Box-Behnken model of response surface methodology (RSM) was employed to optimize B5 medium for the growth of P. oleracea L. hairy root line. According to the results, modelling and optimization conditions, including sucrose, CaCl2.H2O, H2PO4 and NO3-/NH4+ concentrations on maximum dry weight (0.155 g) and noradrenaline content (0.36 mg.g-1 DW) was predicted. These optimal conditions predicted by RSM were confirmed the enhancement of noradrenaline production as an application potential for production by hairy root cultures.

scholarly journals Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

2021 ◽  
Vol 22 (11) ◽  
pp. 5671
Author(s):  
Mohsen Hesami ◽  
Austin Baiton ◽  
Milad Alizadeh ◽  
Marco Pepe ◽  
Davoud Torkamaneh ◽  
...  
Keyword(s):  
Plant Regeneration ◽  
Genetic Engineering ◽  
Suspension Culture ◽  
Secondary Metabolite ◽  
Hairy Root ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Gene Transformation ◽  
Secondary Metabolite Production ◽  
Metabolite Production

For a long time, Cannabis sativa has been used for therapeutic and industrial purposes. Due to its increasing demand in medicine, recreation, and industry, there is a dire need to apply new biotechnological tools to introduce new genotypes with desirable traits and enhanced secondary metabolite production. Micropropagation, conservation, cell suspension culture, hairy root culture, polyploidy manipulation, and Agrobacterium-mediated gene transformation have been studied and used in cannabis. However, some obstacles such as the low rate of transgenic plant regeneration and low efficiency of secondary metabolite production in hairy root culture and cell suspension culture have restricted the application of these approaches in cannabis. In the current review, in vitro culture and genetic engineering methods in cannabis along with other promising techniques such as morphogenic genes, new computational approaches, clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR/Cas9-equipped Agrobacterium-mediated genome editing, and hairy root culture, that can help improve gene transformation and plant regeneration, as well as enhance secondary metabolite production, have been highlighted and discussed.

A reliable and efficient protocol for induction of hairy roots in Agastache foeniculum

Biologia ◽  
2014 ◽  
Vol 69 (7) ◽  
Author(s):  
Elnaz Nourozi ◽  
Bahman Hosseini ◽  
Abbas Hassani
Keyword(s):  
Salicylic Acid ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Performance ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Biochemical Properties ◽  
Separate Experiment ◽  
Transformed Roots

AbstractHairy root culture system is a valuable tool to study the characteristics of gene expression, gene function, root biology, biochemical properties and biosynthesis pathways of secondary metabolites. In the present study, hairy roots were established in Anise hyssop (Agastache foeniculum) via Agrobacterium rhizogenes. Three strains of Agrobacterium rhizogenes (A4, A7 and 9435), were used for induction of hairy roots in four various explants (hypocotyl, cotyledon, one-month-old leaf and five-month-old leaf) of Anise hyssop. The highest frequency of transformation was achieved using A4 strain in one-month-old leaves (51.1%). The transgenic states of hairy root lines were confirmed by PCR (Polymerase chain reaction) method. High performance liquid chromatography analysis revealed that the production of rosmarinic acid (RA) in transformed roots of A. foeniculum was almost 4-fold higher than that of the non-transformed roots. In a separate experiment, hairy roots obtained from one-month-old leaves inoculated with A4 strain, were grown in liquid medium and the effects of different concentrations of salicylic acid (0.0, 0.01, 0.1 and 1 mM) and chitosan (0, 50, 100 and 150 mg L−1) (as elicitor) and sucrose (20, 30, 40 and 50 g L−1) on the growth of hairy roots were evaluated. The results showed that, 30 g L−1 sucrose and 100 mg L−1 chitosan increased the biomass of hairy root cultures and application of salicylic acid reduced the growth of hairy roots compared with control roots.

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