جستجوی مقالات مرتبط با کلیدواژه « Hairy root » در نشریات گروه « پزشکی »

Datura innoxia Mill. is one of the medicinal plant which can produce tropan alkaloids such as hyoscyamine and scopolamine with analgesic and antiasthmatic activities.

In this research, the effect of various factors such as culture media and bacteria strain were studied on hairy roots induction and secondary metabolites production in Datura innoxia.

Strains A4, A13, R1000, MSU, 15834, 2656 and 11325 of Agrobacterium rhizogenes with two different concentrations (OD = 0.5, 1) used for co-cultivation of leaf and stem explants prepared from 1-3-month-old and 5-7-month-old plants to optimize the production of hairy roots and then 6 different media were used for establishment and growth of hairy roots.

The results of data analysis showed that A4 strain with 86.6 % hairy root production was the best strain than others. Also, dark conditions, younger explants and higher concentration of bacteria caused the highest amount of induction of hairy roots. ½ MS medium was recognized as the best medium for the growth of hairy roots. The results of HPLC analysis also showed that the level of hyoscyamine in hairy roots was lower than that of normal roots which, of course, can be compensated by the higher growth of hairy roots.  

The appearance and growth of hairy roots depends on various factors such as bacterial strain, culture medium, explant and its age and other factors that need to be optimized at the beginning of any research.

Citrullus colocynthis is considered as a precious herb due to its medicinal and nutritional values and also for its ability to produce valuable bioactive compounds such as cucurbitacin E and quercetin. The hairy root systems are valuable tools for scaling-up secondary metabolites and for introducing new beneficial traits into herbs. The present research has aimed to develop a protocol for hairy root culture of C. colocynthis using Agrobacterium rhizogenes. After the establishment of the hairy root system, factors such as explant type, bacterial strain, pre-culture period, co-cultivation period, and the use of acetosyringone that affect the efficient transformation of the herb were optimized. Four A. rhizogenes strains (MSU440, A4, A13 and ATCC 15834) and three types of explant (leaf, excised shoot and hypocotyl) were tested. Furthermore, the insertion of transgene into the genome of C. colocynthis was confirmed by polymerase chain reaction analysis. The highest transformation frequency was obtained after the infection of excised shoots by MSU440. Co-cultivation for 48 hours resulted in enhanced transformation frequency, while the results of this research showed that the protocol is better not to include the pre-culturing step. In addition, the presence of acetosyringone in bacterial culture and co-cultivation medium significantly increased the success of C. colocynthis transformation. This study describes an efficient protocol for hairy roots culture of C. colocynthis which can be used for scaling-up the plant active phytochemicals or for genetic manipulations of the plant.

Natural-based drugs are the important bioactive substances that have been used for prevention and treatment of diseases. Natural products should be prepared in commercial scale from relevant medicinal plants. Hence, large amounts of the plants have been needed for extraction and isolation of compounds of natural origin. Plant cells technology is the best strategy for the production of the plant-derived drugs, which have difficulty in high scale preparation. This study was conducted for types, frequencies and efficacies of production methods for natural-based drugs in plant cell technology as alternative method to whole herb. Pharmaceutical and biomedical databases including PubMed/Medline, Scopus, Web of Science, Embase, ProQuest and Google Scholar were searched in this study. Moreover, keywords words were ''secondary metabolite production'', ''pharmaceutical natural compounds'', ''high scale production'', ''cell suspension'', ''immobilized plant cell'', “hairy root”, ''elicitor'', ''substrate'', ''plant cell'', ''callus'', ''medicinal plants'', ''isolation and purification''. The correlations have been investigated by random effect model in an Excel program. Findings of this meta-analysis study showed all production methods had high efficacies and percentages of high scale production from 90 to 100%, which were comparable with conventional direct extractions. In addition to, median efficacy values for cell suspension, callus, hairy root and immobilized plant cell methods in production of selected drugs (atropine, paclitaxel, vincristine, camptothecin and colchicine) with 1124, 257, 797 and 969 events were 92.49 (CI95%: 89.78-95.86), 91.98 (CI95%: 89.13-95.25), 95.69 (CI95%: 92.84-98.68) and 93.86% (CI95%: 91.12-96.35), respectively. The plant cell technology for production of secondary metabolites has various advantages including high accuracy, repeatability and productivity, that is a best strategy for production of natural-based drugs.

Biotechnologic methods are common for secondary metabolites production from the plants and other sources in pharmaceutical sciences. Hairy root cell lines as the biotechnologic method have been used for in vitro production of major plant metabolites. In this study, hairy roots of Hypericum Perforatum have been prepared using the seeds and bio transformed by bacteria. Finally, the hypericin have been producted by the hairy roots. First, the seeds have been incubated in the plant media to hairy roots produced. Then, the hairy roots have been dipped in Rhizobium rizogenes suspension for biotransformation of bacterial genes. Morphological and phytochemical features of hairy roots have been determined in order to select the H. perforatum genotypes with higher hypericin contents. The fresh and dry weight of ten lines clones were measured after 30 days. In addition to, methanolic extracts of final hairy roots have been prepared and hypericin has been isolated and assayed by High Performance Liquid Chromatography. According to the observations, one of ten hairy root lines shows 75-fold higher hypericin content (339.27 ppm) compared to non-transformed H. perforatum (4.58 ppm) in the same of body weight of roots. The clone with the most content of hypericin had significant development of biomass of hairy roots and increase the hypericin production. Transformed clones were varied in morphology, growth, and metabolite productivity. The mentioned methods induce the production of hairy root secondary metabolites in high scale to improve the quality and the quantity of pharmaceutical compounds.

Cosmos bipinnatus is an important medicinal plant with antioxidative, antigenotoxic, anti-inflammatory, and anti-proliferative effects on several cancer cell lines. It has great potential for development as a promising cancer chemo-preventive agent. Hairy root-based culture technology is a new sustainable production platform for producing specific pharmaceutical secondary metabolites.
The current study developed and introduced a reliable transformation system for C. bipinnatus by optimization of aspects important in transformation frequency using Agrobacterium rhizogenes.
Five bacterial strains, including ATCC 15834, ATCC 31798, A7, MAFF-02-10266, and MSU440, 2 explant types (leaf and stem), and 2 co-cultivation media (full MS and ½ MS) were examined. Genomic DNA was extracted using a modified CTAB protocol from putative transgenic root lines and the control root. Transgenic hairy root lines were approved by means of Polymerase Chain Reaction (PCR) using specific rolB gene primers.
The highest ratio of genetically transformed root induction was found from leaf explants using A. rhisogenes strains ATCC15834 and MSU440 (72% to 73%). When ½ MS medium was used as a co-cultivation medium, a significant increase in transformation frequency (84%) was observed.
The MSU440 Agrobacterium strain and ½ MS co-cultivation medium could significantly improve genetic transformation efficiency for establishment of hairy root-based cultures for C. bipinnatus.

The production and purification cost of a recombinant protein in plants is lower than other conventional systems, such as bacteria. Comparing different parts of the plant, the hairy root is known to be a suitable bioreactor to produce recombinant proteins including vaccine candidate immunogens. Among the most important causes of diarrhea, E. coli and Vibrio cholerae cause the disease by producing a toxin. The B subunit of this toxin is an appropriate candidate for vaccine development because of its non-toxicity and exposure to the immune system. To investigate the efficiency of transgenic hairy roots in the production of recombinant protein, two methods were considered: direct (induction of hairy root with recombinant Agrobacterium rhizogenes) and indirect (production of transgenic plant with Agrobacterium tumefaciens and then polluting it with non-transgenic A. rhizogenes). To compare the 2 methods, GUS protein expression was used as a reporter and chimeric protein LSC (consisting of ltB-stxB-ctxB) as an antigenic protein. Transformation of tobacco hairy root was confirmed by genomic Polymerase chain reaction (PCR). Gene expression comparison was investigated by semi-quantitative ELISA and enzymatic activity. The results show that the activity of GUS reporter enzyme in the indirect method is seven times more than that of the direct method. Likewise, the expression of LSC recombinant protein in the indirect transformation was 1.5 times more than in direct method. Comparing of these two methods indicated that the hairy roots in the indirect method yield higher protein content than in the direct method.

Hyoscyamus reticulatus contains two distinguished tropane alkaloids, hyoscyamineand scopolamine and both of the compounds possesspotential acute or chronic toxicity. In the present study, a simple and efficient transformation system was established for in vitro hairy roots induction in Hyoscyamus reticulatus.

Effect of different factors including Agrobacterium rhizogenes strains (A7, 15834, A13 and D7), various explant types (cotyledon, hypocotyl, two weeks old leaf, four weeks old leaf, two weeks old internode and four weeks old internode), two inoculation methods (immersion and injection) and four types of culture media (MS, ½ MS, ¼ MS and B5) on hairy roots induction efficiency in Hyoscyamus reticulatus were tested. In the second part of the experiments, elicitations with different concentrations of colchicine (0, 0.01, 0.03 and 0.05% w/v) and different UV-B exposure time (0, 3, 6 and 9 min) were used to analyze hyoscyamineand scopolamine production.  Transgenic status of hairy roots was confirmed by PCR using specific primers of the rolB gene. The total antioxidant activity was evaluated by DPPH) method.

Induction of hairy roots in H. reticulatus was affected by bacterial strain and explant type. A7 strain and cotyledon explants were detected as the best strain and explants for induction of hairy root in H. reticulatus. Hairy roots growth was significantly affected by medium type. The highest fresh weight was produced in MS and B5 medium.Fresh and dry weight of hairy root reached 1.44 and 0.134 mg at 0.05 percentage of colchicine after 48 h, respectively but in UV-B treatment fresh weight was decreased. In addition, antioxidant activity of hairy root samples treated with colchicine and UV-B increased to 27% (0.05 colchicine) and 26% (UV-B 9 min), respectively compared to the antioxidant activity level in non-transgenic roots (12%) and transgenic roots (18%). The highest amount of hyoscyamine and scopolamine (0.58% and 1.9 %) found in elicited hairy root cultures was 3.2 and 5.1 folds higher than the non-transformed roots (0.18% and 0.37%), respectively. B5 and MS medium were detected as the best appropriate medium for growth of H. reticulatus hairy roots. Antioxidant activity in elicited hairy roots with elicitors increased in comparison to the antioxidant activity level in transgenic and non-transgenic hairy roots.

Hairy root lines developed and elicited in this study can be used to investigate the production of pharmaceutically important metabolites of H. reticulatus.

The seed extract of Silybum marianum contains seven flavonolignans known collectively as silymarin. These metabolites can be produced in hairy root cultures of S. marianum. The effect of different physical factors can change root biomass and silymarin production which has been investigated in the present study.

The effect of different physical factors of temperature (30 ºC/25 ºC, 25 ºC/25 ºC and 15 ºC/20 ºC in 16 h/8 h cycle) and pH (5, 5.7, 6 and 7) were evaluated with respect to the root biomass and silymarin production in hairy root cultures of the plant.

Incubation temperature, 25 ºC /25 ºC promoted the silymarin production in 4-week old hairy roots (0.18 mg/g DW) as compared with the cultures treated with 15 ºC/20 ºC and 30 ºC/25 ºC (0.13 and 0.12 mg/g DW, respectively). Maximal increases in biomass and silymarin accumulation occurred in the root cultures grown in pH 5 and 25 ºC/25 ºC (0.45 g and 0.26 mg/g DW). The content of silybin, isosilybin, silychristin, silydianin were 0.025, 0.024, 0.061 and 0.095 mg/g DW, respectively which were higher than those grown in higher pH.

The results of the present study suggest that 25 ºC/25 ºC and acidic environment of medium are beneficial for silymarin production using hairy root cultures. Furthermore, lipoxygenase (LOX) activity was strongly affected by pH which suggested that acidic environment may act as inducing signal for LOX activity and subsequently greater silymarin production.