Strategies to Improve the Production of Forskolin from Hairy Root Cultures of Coleus forskohlii Briq.

2012 ◽  
Vol 5 (2) ◽  
pp. 1720-1726
Author(s):  
Veeresham C ◽  
C.S. Reddy ◽  
Praveena Ch
Keyword(s):  
Methyl Jasmonate ◽  
Hairy Root ◽  
Casein Hydrolysate ◽  
Leaf Explants ◽  
Basal Medium ◽  
Root Culture ◽  
Suspension Cultures ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Coleus Forskohlii

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.

scholarly journals Effects of methyl jasmonate on the growth and triterpenoid production of diploid and tetraploid Centella asiatica (L.) Urb. hairy root cultures

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Khoa Van Nguyen ◽  
Benyakan Pongkitwitoon ◽  
Thanika Pathomwichaiwat ◽  
Unchera Viboonjun ◽  
Sompop Prathanturarug
Keyword(s):  
Methyl Jasmonate ◽  
Hairy Root ◽  
Hairy Roots ◽  
Centella Asiatica ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Asiatic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Meja Treatment

AbstractIn this study, the effects of methyl jasmonate (MeJA) on the phytomass and triterpenoid production of diploid and tetraploid Centella asiatica hairy roots were investigated. Hairy root cultures were obtained from diploid and induced tetraploid plants of C. asiatica infected by Agrobacterium rhizogenes strain ATCC 43057. MeJA triggered triterpenoid production in both ploidy hairy roots, whereas triterpenoids were not produced in the untreated hairy roots. Among the treatments, the 50 µM MeJA treatment yielded the maximum triterpenoid production in diploid hairy roots of 27.25 ± 0.27 µg/mg Dry weight (DW) total triterpenoid at day 21. For the tetraploid hairy root cultures, the 28th-day hairy root culture produced a maximum amount of triterpenoids of 16.29 ± 6.32 µg/mg DW in response to the 50 µM MeJA treatment, whereas the 100 µM MeJA treatment produced a similar triterpenoid amount (16.31 ± 9.24 µg/mg DW) at day 14. Moreover, in response to 50 µM MeJA, we obtained different ratios of aglycone to glycoside, i.e., 1:7 and 1:2, between the diploid and tetraploid hairy root cultures. Asiaticoside was the dominant phytochemical, followed by asiatic acid and madecassic acid. This study provides valuable information for producing triterpenoids for C. asiatica commercial products and preparations by using hairy root cultures.

Effects of Culture Conditions on Isoflavonoid Levels of Transformed and Non-Transformed Cultures of Lupinus -a Comparison of Suspension and Hairy Root Cultures

1991 ◽  
Vol 46 (9-10) ◽  
pp. 735-742 ◽  
Author(s):  
J. Berlin ◽  
C. Rügenhagen ◽  
M. Rippert ◽  
S. Erdogan
Keyword(s):  
Suspension Culture ◽  
Hairy Root ◽  
Root Culture ◽  
Culture Conditions ◽  
Suspension Cultures ◽  
Stress Factors ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Free Medium ◽  
Lupinus Polyphyllus

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.

scholarly journals Methyl jasmonate enhances ursolic, oleanolic and rosmarinic acid production and sucrose induced biomass accumulation, in hairy roots of Lepechinia caulescens

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11279
Author(s):  
Victor M. Vergara-Martínez ◽  
Samuel E. Estrada-Soto ◽  
Susana Valencia-Díaz ◽  
Karlina Garcia-Sosa ◽  
Luis Manuel Peña-Rodríguez ◽  
...  
Keyword(s):  
Methyl Jasmonate ◽  
Bioactive Compounds ◽  
Hairy Root ◽  
Hairy Roots ◽  
Leaf Explants ◽  
Biomass Accumulation ◽  
Wild Plants ◽  
Bioactive Metabolites ◽  
Hairy Root Cultures ◽  
Root Cultures

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.

Biotransformation of low-molecular-weight alcohols by Coleus forskohlii hairy root cultures

2003 ◽  
Vol 338 (8) ◽  
pp. 729-731 ◽  
Author(s):  
Wei Li ◽  
Kazuo Koike ◽  
Yoshihisa Asada ◽  
Takafumi Yoshikawa ◽  
Tamotsu Nikaido
Keyword(s):  
Molecular Weight ◽  
Hairy Root ◽  
Low Molecular Weight ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Coleus Forskohlii

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.

scholarly journals Enhanced production of valtrate in hairy root cultures of Valeriana jatamansi Jones by methyl jasmonate, jasmonic acid and salicylic acid elicitors

2020 ◽  
Vol 48 (2) ◽  
pp. 839-848
Author(s):  
Shuang ZHAO ◽  
Hong TANG
Keyword(s):  
Salicylic Acid ◽  
Jasmonic Acid ◽  
Methyl Jasmonate ◽  
Hairy Root ◽  
Hairy Roots ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Potent Inducer ◽  
Valeriana Jatamansi ◽  
Pharmacologically Active

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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