scholarly journals Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.

PLoS ONE ◽  
2017 ◽  
Vol 12 (8) ◽  
pp. e0182367 ◽  
Author(s):  
Poornima Devi Gurusamy ◽  
Holger Schäfer ◽  
Siva Ramamoorthy ◽  
Michael Wink
2019 ◽  
Vol 9 (3) ◽  
pp. 415-420
Author(s):  
L. P. Khlebova ◽  
E. S. Brovko ◽  
O. V. Bychkova ◽  
N. V. Pavlova

The hairy root cultures are promising sources of secondary metabolites of plants, including rare and endangered species. They possess genetic and biochemical stability, unlimited growth rate in free-hormone medium, short doubling times, high biosynthetic activity and ecological purity of plant raw materials. The hairy root cultures of Tagetes patula L. can be used to produce biologically active substances with biocidal activity. The study aimed to determine the virulent strain of Agrobacterium rhizogenes and the most effective period of co-cultivation of T. patula leaf explants with an agrobacterium to induce actively growing hairy root cultures. We used 3 strains (A-4b, 8196RT and 15834). The time of infection ranged from 3 to 33 hours in increments of 3 hours. We found that 24 h is the best time of infection to induce hairy roots with the highest transformation efficiency (92%). The wild strain A. rhizogenes 15834 turned out to be the most virulent when infected leaf explants of spreading marigold. This strain provided the maximum transformation effect, reaching 85.4%. We have identified 5 actively growing clones of hairy roots with intensive branching, the growth indices of which were 64-75. In the future, they will be transferred to a liquid medium for biomass accumulation and scaling.


Molecules ◽  
2020 ◽  
Vol 25 (9) ◽  
pp. 2262 ◽  
Author(s):  
Ewa Kochan ◽  
Adriana Nowak ◽  
Małgorzata Zakłos-Szyda ◽  
Daria Szczuka ◽  
Grażyna Szymańska ◽  
...  

American ginseng, Panax quinquefolium (L.), is traditionally used in folk medicine. It exhibits a range of anti-inflammatory, hepatoprotective, anti-diabetic, anti-obesity, anti-hyperlipidemic and anti-carcinogenic effects. Its main components are ginsenosides, also known as panaxosides or triterpene saponins. In order to obtain high yields of ginsenosides, different methods of controlled production are involved, i.e., with hairy root cultures. However, they are still employed under in vitro conditions. Our studies revealed that hairy root cultures subjected to an elicitation process can be considered as a potent source of ginsenosides. The present study examines the biological activity of ginseng hairy root cultures against the Caco-2 human adenocarcinoma cell line. Among our six different clones of P. quinquefolium hairy roots, extracts B and Be (treated with elicitor) were the strongest inhibitors of the cellular metabolic activity. While all extracts induced DNA damage, B and Be also generated reactive oxygen species (ROS) in a concentration-dependent manner, which was correlated with the depletion of the mitochondrial membrane potential and induction of apoptosis. These findings indicate that further research concerning P. quinquefolium hairy root cultures should focus on the activity of rare ginsenosides and other biologically active compound profiles (i.e., phenolic compounds).


2009 ◽  
Vol 64 (11-12) ◽  
pp. 869-874 ◽  
Author(s):  
Rawia Zayed ◽  
Michael Wink

Hairy root cultures of Nicotiana tabacum were set up by excised root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analyzing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of pyridine alkaloids, mainly nicotine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid, in order to stimulate the biosynthesis of pyridine alkaloids. Profile and amounts of pyridine alkaloids were analyzed using capillary GLC-MS. Treatment of the cultures with methyljasmonate (50 μM) increased the alkaloid accumulation ca. 7-fold up to a level of 0.58 mg/g fresh weight as compared to untreated controls. Quercetin (200 μM) enhanced the alkaloid production ca. 4-fold (0.34 mg/g fresh weight) within 24 h. In contrast salicylic acid in all tested concentrations decreased the alkaloid level to 1 μg/g fresh weight. Also the inhibitory effect of salicylic acid on the elicitation effect of methyljasmonate and quercetin was investigated.


2020 ◽  
pp. 209-221
Author(s):  
Elena Valerievna Ambros ◽  
Ol'ga Viktorovna Kotsupiy ◽  
Tat'yana Abdulkhailovna Kukushkina ◽  
Tat'yana Vital'yevna Zheleznichenko ◽  
Tat'yana Ivanovna Novikova

Agrobacterium rhizogenes – mediated genetic transformation of medicinal plant A. penduliflorus Lam. using A4-RT, R-1601, 15834 SWISS strains was performed. The competences for transformation of three types of explants: hypocotyls, cotyledons, and primary shoots were tested. The virulent strain (15834 SWISS) and types of explants for transformation (primary shoots and cotyledons) with high growth index (I) were determined. The frequency of transformation of cotyledons by strain 15834 SWISS after 4 weeks of cultivation was 15.4% (I = 59.6), hypocotyls – 9.1% (I = 7.3) and primary shoots – 37.5% (I = 21.0). After 8 weeks of cultivation I increased 4.5 times for primary shoots (I = 94.5 ± 0.20) and cotyledons (I = 265.8 ± 0.35), for cultures from hypocotyls – 5.97 times (I = 43.6 ± 0.30). The roots’ transgenic status and the absence of agrobacterium contamination were confirmed by PCR analysis using rolB-, virC-specific primers. The lines of hairy roots characterized by active increases of biomass with high content of biologically active metabolites were selected, moreover, the content of metabolites in hairy root cultures exceeded their content in the roots of introduced plants. The maximum accumulation of compounds was found in hairy roots obtained from primary shoots (pectins – up 7.8%, protopectins – up 15.3%) and cotyledons (tannins – up 16.1%, triterpenic saponins – up 30.5%) after 8 weeks of cultivation. High perfor-mance liquid chromatography (HPLC) analysis demonstrated that hydrolysates of extracts of hairy roots from primary shoots contained 2 flavonol aglycones – quercetin and isorhamnetin whereas ethanol extracts were characterized by presence of quercetin and 4 flavonoid components. In hairy roots from cotyledons the maximum of phenolic compounds (PСs) content did not differ significantly at 8 and 12 weeks of cultivation (1.38 ± 0.01 and 1.49 ± 0.06% of dry weight, respectively). The content of PCs in hairy roots from primary shoots increased two-fold from 4 to 12 weeks of cultivation (up 1.24 ± 0.18%). To the best of our knowledge, this is the first efficient protocol reported for the establishment of hairy root cultures in A. penduliflorus using A. rhizogenes.


Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2828 ◽  
Author(s):  
Bertóti ◽  
Böszörményi ◽  
Alberti ◽  
Béni ◽  
M-Hamvas ◽  
...  

Horseradish hairy root cultures are suitable plant tissue organs to study the glucosinolate–myrosinase–isothiocyanate system and also to produce the biologically active isothiocyanates and horseradish peroxidase, widely used in molecular biology. Fifty hairy root clones were isolated after Agrobacterium rhizogenes infection of surface sterilized Armoracia rusticana petioles and leaf blades, from which 21 were viable after antibiotic treatment. Biomass properties (e.g. dry weight %, daily growth index), glucosinolate content (analyzed by liquid chromatography-electronspray ionization-mass spectrometry (LC-ESI-MS/MS)), isothiocyanate and nitrile content (analyzed by gas chromatography-mass spectrometry (GC-MS)), myrosinase (on-gel detection) and horseradish peroxidase enzyme patterns (on-gel detection and spectrophotometry), and morphological features were examined with multi-variable statistical analysis. In addition to the several positive and negative correlations, the most outstanding phenomenon was many parameters of the hairy root clones showed dependence on the organ of origin. Among others, the daily growth index, sinigrin, glucobrassicin, 3-phenylpropionitrile, indole-3-acetonitrile and horseradish peroxidase values showed significantly higher levels in horseradish hairy root cultures initiated from leaf blades.


Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
T Hasanloo ◽  
H Rahnama ◽  
R Sepehrifar ◽  
MR Shams

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