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1.
The study assesses the influence of different concentrations of nitrogen and phosphorus sources on ginsenoside biosynthesis in Panax quinquefolium hairy roots cultivated in shake flasks and a nutrient sprinkle bioreactor. The saponin content was determined using HPLC. The maximum yield (12.45 mg g?1 dw) of the sum of six examined ginsenosides (Rb1, Rb2, Rc, Rd, Re and Rg1) in hairy roots cultivated in shake flasks was achieved in modified Gamborg B-5 medium containing 0.83 mM l?1 phosphate, 12.4 mM l?1 nitrate and 0.5 mM l?1 ammonium. The yield itself was 1.93 times higher than that achieved in standard Gamborg medium. The modified medium also favourably influenced the biosynthesis of studied saponins in bioreactor cultures. The saponin content (35.11 mg g?1 d.w.) was 2.75-times higher than that achieved in control medium.  相似文献   

2.

Microshoots of the East Asian medicinal plant species Schisandra chinensis (Chinese magnolia vine) were grown in bioreactors characterized by different construction and cultivation mode. The tested systems included two continuous immersion systems—a cone-type bioreactor (CNB) and a cylindric tube bioreactor (CTB), a nutrient sprinkle bioreactor (NSB), and two temporary immersion systems (TIS)—RITA® and Plantform. Microshoots were grown for 30 and 60 days in the MS medium enriched with 1 mg l?1 NAA and 3 mg l?1 BA. The accumulation of two groups of phenolic compounds: phenolic acids and flavonoids in the bioreactor-grown S. chinensis biomass, was evaluated for the first time. In the microshoot extracts, seven phenolic acids: chlorogenic, gallic, p–hydroxybenzoic, protocatechuic, syringic, salicylic and vanillic, and three flavonoids: kaempferol, quercitrin and rutoside, were identified. The highest total amount of phenolic acids (46.68 mg 100 g?1 DW) was recorded in the biomass maintained in the CNB for 30 days. The highest total content of flavonoids (29.02 mg 100 g?1 DW) was found in the microshoots maintained in the NSB for 30 days. The predominant metabolites in all the tested systems were: gallic acid (up to 10.01 mg 100 g?1 DW), protocatechuic acid (maximal concentration 16.30 mg 100 g?1 DW), and quercitrin (highest content 21.00 mg 100 g?1 DW).

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3.
Hairy roots of Dracocephalum moldavica L. were induced using Agrobacterium rhizogenes strain A4. Transformed roots were obtained from shoot explants with low transformation frequency of up to 3 %. The effects of different liquid media: Murashige and Skoog (MS), Gamborg et al. (B5) and Woody Plant (WP) with full- and half-strength (½MS, ½B5, ½WP), on biomass accumulation and rosmarinic acid (RA) content were investigated. The hairy roots were cultured in photoperiod (16 h light/8 h dark) and darkness. Biomass of D. moldavica hairy roots was the highest (7.23 g flask?1 of fresh weight and 0.89 g flask?1 of dry weight) in the cultures grown in WP medium under periodic light. Ultra performance liquid chromatography analysis revealed the highest RA content (78 mg g?1 dry wt) in roots cultured in ½B5 medium under photoperiod conditions. It was about tenfold higher compared to roots of field-grown mother plants. Antioxidant activities and total phenolic contents of methanolic extracts of D. moldavica hairy roots cultured in ½B5 and WP media under photoperiod and darkness and roots of field grown plants were compared. All extracts were investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging and phosphomolybdenum reduction assays. Total phenolic contents were estimated by the Folin–Ciocalteu method. The methanolic extract of D. moldavica hairy roots grown in ½B5 medium under photoperiod possessed the strongest effects on reducing Mo and DPPH radical scavenging. The activities were significantly higher (p ≤ 0.05) than those of methanolic extract of roots of intact plants grown in the field. The most active methanolic extract of hairy roots was characterized by the highest level of rosmarinic acid and total content of phenolic compounds.  相似文献   

4.
Hairy root cultures of Salvia sclarea were grown in shake flasks and 10 L nutrient sprinkle bioreactor, running for 30 days and the effects of methyl jasmonate (MJ) on their growth and capacity to accumulate diterpenoids were measured. We found that MJ concentration and exposure time to the elicitor were factors that strongly affected the diterpenoid production. The highest diterpenoid accumulation (67.5 ± 7.1 mg g−1 dry weight, calculated as a sum of ferruginol, salvipisone, aethiopinone and 1-oxoaethiopinone) without reduction of biomass, was achieved when the 23-day-old hairy roots in bioreactor culture were exposed to 125 μM MJ for 7 days. The roots produced 9 and 3.8 times as much aethiopinone (40 ± 5.9 mg g−1 dry weight) and salvipisone (12.6 ± 0.4 mg g−1 dry weight), respectively, as roots cultured in shake flasks. Our results imply that cultivation of S. sclarea hairy roots in sprinkle bioreactor after elicitation with MJ may be valuable to enhance production of the bioactive diterpenoids.  相似文献   

5.
Lactoalbumin hydrolysate (LH) at 100 mg L−1 with methyl jasmonate (MJ) at 2 mg L−1 synergistically stimulated ginsenoside accumulation in Panax quinquefolium cells compared with 100 mg L−1 LH. Combination elicitors led to higher ginsenoside productivity (45.93 mg L−1) than single treatment of 100 mg L−1 LH (31.37 mg L−1). This present result will be helpful in providing a tool for enhancing the productivity of ginsenoside by Panax quinquefolium cell cultures on a commercial scale.  相似文献   

6.
The study assessed the influence of sugar concentration (10, 20, 30, 50, 70, 100, 120 g l?1) on growth and ginsenoside biosynthesis in Panax quinquefolium hairy roots cultivated in shake flasks and a nutrient sprinkle bioreactor. The highest growth rate was achieved in medium containing 3–5 % sucrose. More than 70 g l?1 or less than 20 g l?1 sugar content in the medium induces significant inhibition of root growth when cultivated in shake flasks. The saponin content was determined using HPLC. The maximum yield (above 9 mg g?1 d.w.) of the sum of six examined ginsenosides (Rb1, Rb2, Rc, Rd, Re and Rg1) in hairy roots cultivated in shake flasks was obtained with 30 g l?1 sucrose in the medium. The sucrose concentration in the medium was found to correlate with saponin content in bioreactor-cultured specimens. A higher level of protopanaxadiol derivatives was found for lower (20 and 30 g l?1) sucrose concentrations; higher sucrose concentrations (50 and 70 g l?1) in the medium stimulated a higher level of Rg group saponins.  相似文献   

7.
In this article, ginsenosides and polysaccharide contents in suspension cells and native roots of Panax quinquefolium L. were studied. In order to enhance the contents of ginsenosides and polysaccharide in P. quinquefolium suspension cells, we tested the effects of lactoalbumin hydrolysate on the growth of P. quinquefolium suspension cell, synthesis of ginsenosides and polysaccharide in flask and bioreactor. In flask culture, cells growth ratio was significantly enhanced by the addition of lower concentration of lactoalbumin hydrolysate. Addition of 100 mg L−1 lactoalbumin hydrolysate significantly enhanced the contents of total saponins (5.44 mg g−1 DW) and the contents were 3.89-fold over the control group. Addition of lactoalbumin hydrolysate significantly promoted the accumulation of polysaccharide, except 200 mg L−1 lactoalbumin hydrolysate. The highest total saponins yield (36.72 mg L−1 DW) and polysaccharide yield (0.83 g L−1 DW) were obtained at 100 mg L−1 lactoalbumin hydrolysate. In a 5-L stirred tank bioreactor, the highest contents of total saponins and TRb group ginsenosides were achieved on day 26, while the effect of lactoalbumin hydrolysate on the contents of TRg group ginsenosides were insignificant. This result suggests that lactoalbumin hydrolysate might have triggered the enzyme activities for the synthesis of TRb group ginsenosides. Overall, the highest total saponins yield (31.37 mg L−1 DW) and polysaccharide yield (1.618 g L−1 DW) were obtained on day 26 and day 24 respectively and the polysaccharide yield was 1.95-fold higher than the shake flask culture (0.83 g L−1 DW). These results provided theoretical reference for two-stage culture in suspension cells of P. quinquefolium in bioreactor.  相似文献   

8.
Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension cultures or on agar solidified medium. Psoralen content was ~3 mg g−1 DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious roots.  相似文献   

9.
Rehmannia glutinosa Libosch., a valuable medicinal plant, was successfully propagated in vitro using shoot tip explants. Shoot multiplication was performed in glass tubes and in a nutrient sprinkle bioreactor. A mixture of 0.1 mg L?1 indole-3-acetic acid (IAA) and 1.0 mg L?1 of 6-benzylaminopurine in Murashige and Skoog (MS) agar-solidified medium proved the best combination for multiple shoot induction, yielding 8.2 shoots per explant after 4 weeks of culture in glass tubes. The number of shoots increased to 21 per explant when the same combination of growth regulators was used in a nutrient sprinkle bioreactor. The shoots rooted with a frequency of 93 % after 6 weeks of culture on MS agar medium supplemented with IAA (0.1 mg L?1) before being acclimatized in the greenhouse. The antioxidant activities of methanolic extracts from the leaves and roots of the in vitro-regenerated plants of R. glutinosa cultivated in the greenhouse were evaluated using four in vitro assays: scavenging of free radicals (DPPH and ABTS), transition metal reduction and total antioxidant activity phosphomolybdenum test. In all cases, the methanolic extract from leaves demonstrated better antioxidant activity than those taken from roots. A strong correlation was found between total phenolic and flavonoid content, and the antioxidant capacity of the studied extracts.  相似文献   

10.
Leaves and flowers from Tunisian Globularia alypum were assessed for their phenolic and flavonoid contents and antioxidant activity. Phenolic (15.5–22.30 mg GAE g?1 DW) and flavonoid (3.63–4.72 mg RE g?1 DW) contents varied according to plant part. Globularia alypum leaves exhibited the highest phenol and flavonoid contents, and displayed the highest antioxidant activity, based on radical‐scavenging activity (295 μmol TEAC g?1 DW) and ferric reducing power (24.27 mmol Fe2+/l). The obtained results demonstrated that some of the isolated compounds play an important role for the antioxidant activity of G. alypum and give a scientific basis to the use of this plant in traditional medicine. The methanolic extract of G. alypum could be considered as a source of potential antioxidants and will promote the reasonable usage of this plant in food technology and processing as well as for medical use.  相似文献   

11.
Astragalus membranaceus is one of the most widely used traditional medicinal herbs in China, but the time required to generate a useful product in the field production is long. The growth of adventitious root cultures was compared between cultures grown in solid, liquid, or a 5-L balloon-type bubble bioreactor. The maximum growth ratio (final dry weight/initial dry weight) was determined for adventitious roots grown in the bioreactor. Studies carried out to optimize biomass production of adventitious roots compared adventitious root growth from various inoculum root lengths, inoculum densities, and aeration volume in the bioreactors. The maximum growth ratio occurred in treatments with a 1.5-cm inoculum root length, with 30 g (fresh weight) of inoculum per bioreactor or with an aeration volume of 0.1 vvm (air volume/culture medium volume per min). The polysaccharide, saponin, and flavonoid content of roots from bioreactor-grown cultures were compared to roots from field-grown plants grown for 1 and 3 yr. Total polysaccharide content of adventitious roots in the bioreactor (30.0 mg g−1 dry weight (DW)) was higher than the roots of 1-yr-old (13.8 mg g−1 DW) and 3-yr-old (21.1 mg g−1 DW) plants in the field. Total saponin (3.4 mg g−1 DW) and flavonoid (6.4 mg g−1 DW) contents were nearly identical to 3-yr-old roots and higher than that of 1-yr-old roots under field cultivation.  相似文献   

12.
Microbial resistance and other emerging health risk problems related to the side effects of synthetic drugs are the major factors that result in the research regarding natural products. Fruits, leaves, seeds, and oils-based phyto-constituents are the most important source of pharmaceutical products. Plant extract chemistry depends largely on species, plant components, solvent utilized, and extraction technique. This study was aimed to compare the ethanolic extracts of a mangrove plant, i.e., Avicennia marina (1E: Lower half of A. marina‘s pneumatophores, 2E: A. marina‘s leaves, 3E: Upper half of A. marina‘s pneumatophores, and 4E: A. marina‘s shoots), with non-mangrove plants, i.e., Phragmites australis (5E: P. australis‘s shoot), and Moringa oleifera (6E: M. oleifera‘s leaves) for their antimicrobial activities, total phenolic contents, antioxidant activity, and cytotoxicity potential. The antimicrobial activity assays were performed on gram-positive bacteria (i.e., Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (i.e., Escherichia coli, and Pseudomonas aeruginosa), and fungi (i.e., Aspergillus niger, Candida albicans, and Rhizopus spp.). We estimated antioxidant activity by TAC, DPPH, and FRAP assays, and the cytotoxicity was evaluated by MTT assay. The results of antimicrobial activities revealed that B. subtilis was the most sensitive to the tested plant extracts compared to S. aureus, while it only showed sensitivity to 6E and Imipenem. 5E and 6E showed statistically similar results against P. aeruginosa as compared to Ceftazidime. E. coli was the most resistant bacteria against tested plant extracts. Among the tested plant extracts, maximum inhibition activity was observed by 6E against A. niger (22 ± 0.57 mm), which was statistically similar to the response of 6E against C. albicans and 3E against Rhizopus spp. 2E did not show any activity against tested fungi. We found that 6E (208.54 ± 1.92 mg g?1) contains maximum phenolic contents followed by 1E (159.42 ± 3.22 mg g?1), 5E (131.08 ± 3.10 mg g?1), 4E (i.e., 72.41 ± 2.96 mg g?1), 3E (67.41 ± 1.68 mg g?1), and 2E (48.72 ± 1.71 mg g?1). The results depict a significant positive correlation between the phenolic contents and the antioxidant activities. As a result, phenolic content may be a natural antioxidant source.  相似文献   

13.
Hairy roots of Nasturtium officinale, Barbarea verna and Arabis caucasica with active glucosinolate-myrosinase system were obtained after transformation with Agrobacterium rhizogenes. Hairy roots of N. officinale produced phenylalanine-derived gluconasturtiin and glucotropaeolin (max. 24 and 7 mg g−1 DW). B. verna and A. caucasica hairy roots produced gluconasturtiin (max. 41 mg g−1 DW) and methionine-derived glucoiberverin (max. 32 mg g−1 DW), respectively. Treatment of the roots with amino acid precursors of glucosinolate or/and cysteine biosynthesis increased levels of glucosinolate production, combinations of phenylalanine with cysteine (for gluconasturtiin and glucotropaeolin) and methionine with o-acetylserine (for glucoiberverin) were the most effective.  相似文献   

14.
A genetic transformation method using Agrobacterium rhizogenes was developed for Harpagophytum procumbens. The influence of three factors on hairy root formation was tested: bacterial strains (A4 and ATCC 15834), various types of explants and acetosyringone (AS) (200 μM). The highest frequency of transformation (over 50% of explants forming roots at the infected sites after 6 weeks of culture on Lloyd and McCown (WP) medium) was achieved using a combination of nodal stem explants and A. rhizogenes strain A4. The addition of 200 μM AS to root induction medium was found to enhance hairy root induction but its effect varied depending on bacterial strain and explant type. Three of the most vigorously growing hairy root clones of H. procumbens were chosen and analyzed for accumulation of iridoid and phenylethanoid glycosides. The transgenic nature of these root clones was confirmed by PCR amplification; they were positive for rolB and rolC genes. Harpagoside, verbascoside and isoverbascoside were identified by HPLC and LC–ESI-MS as the major compounds from all analyzed hairy root clones. The Hp-3 root clone showed the higher harpagoside content (0.32 mg g−1 dry wt.) compared with other analyzed transformed and non-tuberized untransformed roots of H. procumbens. However, the level of the compound in the hairy root clone was lower than that detected in a sample of commercially available root tubers of H. procumbens. The Hp-3 root clone also produced high amounts of verbascoside and isoverbascoside (8.12 mg g−1 dry wt. and 9.97 mg g−1 dry wt., respectively) comparable to those found in root tubers.  相似文献   

15.
16.
The biomass and concentration of bioactive quinone methide-type diterpenes in hairy roots of Salvia austriaca were determined and compared with levels of these metabolites in roots of field-grown plants. The cultures were maintained in shake flasks and a nutrient sprinkle bioreactor. Diterpene production was more efficient in the shake flask root culture than the bioreactor one. Biomass and diterpene production within the shake flask culture was evaluated using Schenk and Hildebrandt (SH), Gamborg (B5), and woody plant medium (WPM), with both full- and half-strength macro and micronutrient concentrations (1/2 SH, 1/2 B5, and 1/2 WPM). Among the tested media, SH medium proved to be most effective for biomass and diterpene production. In this medium, the transformed roots accumulated the levels of taxodone (3.89?mg?g?1 DW; equivalent to 63.3?mg?L?1), taxodione (1.15?mg?g?1 DW; equivalent to 17.4?mg?L?1), 15-deoxy-fuerstione (2.15?mg?g?1 DW; equivalent to 32.5?mg?L?1), and 7-(2′-oxohexyl)-taxodione (0.076?mg?g?1 DW; equivalent to 1.1?mg?L?1). Three diterpenes were also detected in the roots of S. austriaca intact plants, but their concentrations were lower than those in hairy root culture. No 7-(2′-oxohexyl)-taxodione was found in the roots of field-grown plants. The hairy roots were able to maintain high metabolite levels even for 6 years of cultivation. Taxodone, taxodione, 15-deoxy-fuerstione, and 7-(2′-oxohexyl)-taxodione were tested for in vitro activity against Trypanosoma brucei rhodesiense, T. cruzi, and Plasmodium falciparum and their cytotoxicity was determined using L6 cells. Among these compounds, taxodione was the most active against T. brucei rhodesiense [IC50?=?0.05?µM with high selectivity, selectivity index (SI)?=?38]. Taxodione was found to inhibit the growth of P. falciparum and T. cruzi by 50% at respective concentrations of 1.9 and 7.1?µM (SI values of 1.0 and 0.27). Other diterpenoids demonstrated weaker activity against tested parasites (IC50 values ranging from 0.62 to 194.7?µM) and lower selectivity (SI value ranged from 0.4 to 5.0).  相似文献   

17.
Erodium cicutarium is known for its total polyphenolic content, but this work reveals the first highly detailed profile of E. cicutarium, obtained with UHPLC‐LTQ OrbiTrap MS4 and UHPLC‐QqQ‐MS/MS techniques. A total of 85 phenolic compounds were identified and 17 constituents were quantified. Overall, 25 new compounds were found, which have not yet been reported for the Erodium genera, or the family Geraniaceae. Along with methanolic extracts, the so far poorly investigated water extracts exhibited in vitro antioxidant activity according to all performed assays, including the ferric reducing/antioxidant power assay (FRAP), 2,2‐diphenyl‐1‐picrylhydrazyl assay (DPPH), 2,2′‐azinobis(3‐ethylbenzthiazoline‐6‐sulfonic acid) assay (ABTS) and cupric ion reducing antioxidant capacity assay (CUPRAC). Elemental composition analysis performed with inductively coupled plasma atomic emission spectrometry (ICP‐AES) and, additionally, hydride generation atomic absorption spectrometry (HydrEA‐ETAAS) showed six most abundant elements to be decreasing as follows: Mg>Ca>K>S>P>Na, and gave first data regarding inorganic arsenic content (109.3–248.4 ng g?1). These results suggest E. cicutarium to be a valuable source of various phenolic compounds with substantial potential for further bioactivity testing.  相似文献   

18.
Phenol is a common pollutant which is found in wastewater of many industries and removal of phenol from the industrial effluents is a major challenge. Recently, the use of hairy roots has been probed for the removal of phenol. In the present study, phenol at various concentrations (100–500 mg L−1) was treated with hairy roots of Helianthus annuus (sunflower hairy roots, SHRs). SHRs removed 100 mg L−1 of phenol after 144 h of incubation. The effect of polyethylene glycol (PEG), l-proline and d-glucose on the rate of phenol removal was also studied. l-proline enhanced the removal efficiency of SHRs resulting in the removal of 100 mg L−1 of phenol after 24 h while PEG did not show any effect on removal. Peroxidase activity was induced after 24 h of phenol addition. Phenol metabolism to generate catechol as a major metabolite was confirmed using HPLC and GC–MS analyses. The detection of small amounts of cis-cis muconic acid and fumaric acid in the reaction medium suggests that these metabolites are produced from the ring cleavage of catechol. The phytotoxicity and cytotoxicity results suggest the non-toxic nature of the resulting phenol metabolites.  相似文献   

19.
A scale-up culture of adventitious roots of ginseng was established using a balloon-type bubble bioreactor (BTBB). Maximum growth rates of ~52-fold and ~50-fold in 3 and 5 L BTBBs were obtained, respectively after 40 days of inoculation, which was significantly higher than that in 0.5 L conical flask (~15-fold). Gradually scale-up culture of adventitious roots increased the root biomass, while the contents of ginsenoside and polysaccharides were not affected. This study also revealed that radical scavenging activity of dried adventitious roots on 1,1-diphenyl-2-picrylhydrazyl was higher than that of native roots at 20–100 mg L−1 methanolic extract.  相似文献   

20.
We investigated the effect of Agrobacterium rhizogenes-mediated transformation on antioxidant activity of Artemisia vulgaris “hairy” roots. It appeared that transformation may increase flavonoid content as well as DPPH-scavenging activity and ability to reduce Fe3+ as compared to the non-transformed plants. Some “hairy” roots accumulated flavonoids up to 73.1?±?10.6?mg RE/g DW (while the amount of flavonoids in the leaves of non-transformed plants was up to 49.4?±?5.0?mg RE/g DW). DPPH-scavenging activity of some “hairy” root lines was 3–3.8 times higher than such one of the roots of the control plants. The Fe3+-reducing power of most transgenic root extracts exceeded such power of the extracts of the roots of the control plants. The decrease in SOD activity was found in the most “hairy” root lines compared to the control roots. The increase of flavonoid content correlated with the increase of ability of extracts to scavenge DPPH*- radical and Fe3+ - reducing power. No correlation between SOD activity of extracts and concentration of flavonoids was found (p?≥?0.2).Thus, transformation has led to the alteration in flavonoid accumulation and antioxidant activity in A. vulgaris “hairy” roots. Transgenic roots with high-antioxidant properties can be selected after A. rhizogenes-mediated transformation.  相似文献   

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