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

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.

2015 ◽  
Vol 10 (11) ◽  
pp. 1934578X1501001
Author(s):  
Yeon Bok Kim ◽  
Darwin W. Reed ◽  
Patrick S. Covello

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.


2008 ◽  
Vol 63 (9-10) ◽  
pp. 691-698 ◽  
Author(s):  
Siriwan Phongprueksapattana ◽  
Waraporn Putalun ◽  
Niwat Keawpradub ◽  
Juraithip Wungsintaweekul

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.


2017 ◽  
Vol 14 (2) ◽  
pp. 673-677 ◽  
Author(s):  
Do Yeon Kwon ◽  
Haeng Hoon Kim ◽  
Jong Seok Park ◽  
Sang Un Park ◽  
Nam Il Park

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.


Molecules ◽  
2019 ◽  
Vol 24 (16) ◽  
pp. 2907 ◽  
Author(s):  
Abdulwadood Shakir Mahmood Alsoufi ◽  
Cezary Pączkowski ◽  
Marek Długosz ◽  
Anna Szakiel

The aim of the study was the evaluation of the efficiency of selected abiotic elicitors, i.e., silver and cadmium ions, ultrasound, and UV-C irradiation, in the stimulation of triterpenoid biosynthesis, accumulation, and saponin secretion in Calendula officinalis hairy root cultures. Apart from the possible enhancement of triterpenoid production, the relationship between primary and secondary metabolism (represented respectively by sterols and pentacyclic triterpenes), modifications of the sterol compositional profile, and fluctuations in the total triterpenoid content were monitored in the performed experiments. The main phenomenon observed as a response to heavy metal treatment was the stimulation (up to 12-fold) of the secretion of saponins, accompanied by significant changes in sterol composition. Ultrasound stimulated the secretion of saponins (up to 11-fold); however, it exerted diverse influences on the triterpenoid content in hairy root tissue (stimulating or decreasing) depending on the duration of the exposure to the elicitor. UV-C radiation caused a slight increase in the content of both sterols and saponins in hairy root tissue, and stimulated saponin secretion up to 8.5-fold. The expected symptoms of the competition between the biosynthetic pathways of sterols and pentacyclic triterpenoids were less evident in reactions to abiotic stressors than those reported previously for biotic elicitors.


2005 ◽  
Vol 3 (2) ◽  
pp. 90-100 ◽  
Author(s):  
Michael Wink ◽  
A. Wilhelm Alfermann ◽  
Rochus Franke ◽  
Bernhard Wetterauer ◽  
Melanie Distl ◽  
...  

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.


2016 ◽  
Vol 11 (9) ◽  
pp. 1934578X1601100 ◽  
Author(s):  
Chang Ha Park ◽  
Aye AyeThwe ◽  
Sun Ju Kim ◽  
Jong Seok Park ◽  
MariadhasValan Arasu ◽  
...  

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.


2014 ◽  
Vol 70 (4) ◽  
pp. 261-265 ◽  
Author(s):  
Agnieszka Pietrosiuk ◽  
Mirosława Furmanowa

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.


Author(s):  
Abdulwadood Shakir Mahmood Alsoufi ◽  
Cezary Pączkowski ◽  
Marek Długosz ◽  
Anna Szakiel

The aim of the study was the evaluation of the efficiency of selected abiotic elicitors, i.e. silver and cadmium ions, ultrasound, and UV-C irradiation, in the stimulation of triterpenoid biosynthesis, accumulation, and saponin secretion in Calendula officinalis hairy root cultures. Apart from the possible enhancement of triterpenoid production, the relationship between primary and secondary metabolism (represented respectively by sterols and pentacyclic triterpenes), modifications of the sterol compositional profile, and fluctuations in the total triterpenoid content were monitored in the performed experiments. The main phenomenon observed as a response to heavy metal treatment was the stimulation (up to 12-fold) of the secretion of saponins, accompanied by significant changes in sterol composition. Ultrasound stimulated the secretion of saponins (up to 11-fold); however, it exerted diverse influences on the triterpenoid content in hairy root tissue (stimulating or decreasing) depending on the duration of the exposure to the elicitor. UV-C radiation caused a slight increase in the content of both sterols and saponins in hairy root tissue, and stimulated saponin secretion up to 8.5-fold. The expected symptoms of the competition between the biosynthetic pathways of sterols and pentacyclic triterpenoids were less evident in reactions to abiotic stressors than those reported previously for biotic elicitors


Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
R Bertóti ◽  
Á Alberti ◽  
A Böszörményi ◽  
R Könye ◽  
T Horváth ◽  
...  

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