scholarly journals Preliminary study on the preservation of Catharanthus roseus (L.) G. Don hairy root lines induced by Agrobacterium rhizogenes isolated in Vietnam

2018 ◽  
Vol 1 (6) ◽  
pp. 86-95
Author(s):  
Nhut Nhu Nguyen ◽  
Le Van Bui
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Suitable Medium ◽  
Transformed Hairy Roots ◽  
Preliminary Study

Agrobacterium rhizogenes-transformed hairy roots from Catharanthus roseus have been widely used in research and in life. Consequently, methods for preservation are essential to maintain valuable hairy root lines. Our results showed that Gamborg’B5 was the most suitable medium for hairy roots from VIN002 and VIN005 while solid White media was more comfortable for the hairy lines from VIN022 and VIN077. All hairy root lines must be preserved in the dark at 25–27 oC. Under suitable conditions, the rate of lines growing normally reached 69.3, 67.0, 57.3 and 60.7 % for VIN002, VIN005, VIN072, and VIN077, respectively. There were 90.0, 93.3, 80.0 and 93.3 % of lines could preserve rolB, a gene which has important effect on the growth of hairy roots.

scholarly journals Study on the growth of Agrobacterium rhizogenes-induced Catharanthus roseus hairy roots in liquid media

2019 ◽  
Vol 2 (1) ◽  
pp. 5-12
Author(s):  
Nguyen Nhu Nhut ◽  
Bui Van Le
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Growth Kinetics ◽  
Exponential Phase ◽  
Liquid Media ◽  
Hairy Root Line ◽  
Initial Ph ◽  
Initial Results

Catharanthus roseus hairy roots induced by Agrobacterium rhizogenes get potential to apply to many fields. The suitable conditions for culturing depend on each hairy root line. In shaken liquid media in dark at 25 oC, VIN002-12 and VIN005-07 hairy root lines had the best growth in hemi-concentrated Gamborg’B5 media while VIN072-15 and VIN077-09 lines showed the best growth in in hemi-concentrated White media. The appropriate initial pH of medium for the lines was in range of 5.7–6.5. The hairy root lines used 2–5% sucrose as an optimal source of carbon for their growth. In selected conditions, the growth kinetics curves showed the end of exponential phase at the 28th day of culture with VIN005-07 and VIN072- 15 lines whereas at the 35th day with VIN002-12 and VIN077-09 lines. The initial results are quite possible to produce biomass for researches on four hairy root lines in the future.  

scholarly journals Isolation and identification of Agrobacterium rhizogenes inducing hairy roots of Catharanthus roseus in vivo

2017 ◽  
Vol 1 (T1) ◽  
pp. 48-59
Author(s):  
Nhut Nhu Nguyen ◽  
Le Van Bui
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Root Formation ◽  
Isolation And Identification ◽  
16S Rrna Sequence ◽  
Pathogenicity Tests ◽  
Foreign Genes

Agrobacterium rhizogenes is a common tool for transforming foreign genes into plant cell and inducing hairy root formation. A total of 274 colonies of Agrobacterium species was isolated on selective MG-Te medium from 235 samples of rhizophere soils. There were only thirteen colonies belonged to A. rhizogenes being able to efficiently induce the hairy root formation of Catharanthus roseus in vivo based on the morphological, biochemical, and pathogenicity tests. These thirteen colonies were identified as A. rhizogenes based on 16S rRNA sequence and possessing rolABC genes. The results indicated A. rhizogenes was universal in rhizophere soil in Vietnam and riched transgenic tools for researching C. roseus hairy roots in the future.

scholarly journals Hairy root induction in Catharanthus roseus by various strains of Agrobacterium rhizogenes isolated in Vietnam

2016 ◽  
Vol 19 (2) ◽  
pp. 44-52 ◽  
Author(s):  
Nhut Nhu Nguyen ◽  
Le Van Bui
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Root Production ◽  
Root Induction ◽  
Promising Tool ◽  
Enhanced Production ◽  
Rolb Gene ◽  
Polymerase Chain

Catharanthus roseus is a well known medicinal plant. It produces several phytocompounds and many of which show anticancerous properties. However the yields of these compounds are very low. Recently, induction of C. roseus hairy roots by Agrobacterium rhizogenes, is interested as a promising tool for the enhanced production of these metabolites. In this research, wounded leaves from four strains of C. roseus were infected with various strains of A. rhizogenes isolated in Vietnam to provide more information about the induction efficiency of hairy roots. In this experiment, after 3 weeks of infection, the presence of rolB gene in hairy roots were analysed by polymerase chain reaction. All of 13 A. rhizogenes strains could induce the formation of hairy root in C. roseus. The A. rhizogenes C18 strain had the highest induction percentage in C. roseus VIN002, VIN005, and VIN072 with 59.4 %, 50.3 %, and 40.0 % respectively. And the same result was ontained at 26.7% by A. Rhizogenes C26 for C. roseus VIN077 rhizogenes strains. This result identified two A. rhizogenes strains C18 and C26 as potential transformation tools for hairy root production from C. roseus.

scholarly journals Preliminary results of indole alkaloids production in different roots of Catharanthus roseus cultured in vitro

2014 ◽  
Vol 70 (4) ◽  
pp. 261-265 ◽  
Author(s):  
Agnieszka Pietrosiuk ◽  
Mirosława Furmanowa
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Catharanthus Roseus ◽  
Indole Alkaloids ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Preliminary Results ◽  
Intact Plants

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.

A reliable and efficient protocol for induction of hairy roots in Agastache foeniculum

Biologia ◽  
2014 ◽  
Vol 69 (7) ◽  
Author(s):  
Elnaz Nourozi ◽  
Bahman Hosseini ◽  
Abbas Hassani
Keyword(s):  
Salicylic Acid ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Performance ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Biochemical Properties ◽  
Separate Experiment ◽  
Transformed Roots

AbstractHairy root culture system is a valuable tool to study the characteristics of gene expression, gene function, root biology, biochemical properties and biosynthesis pathways of secondary metabolites. In the present study, hairy roots were established in Anise hyssop (Agastache foeniculum) via Agrobacterium rhizogenes. Three strains of Agrobacterium rhizogenes (A4, A7 and 9435), were used for induction of hairy roots in four various explants (hypocotyl, cotyledon, one-month-old leaf and five-month-old leaf) of Anise hyssop. The highest frequency of transformation was achieved using A4 strain in one-month-old leaves (51.1%). The transgenic states of hairy root lines were confirmed by PCR (Polymerase chain reaction) method. High performance liquid chromatography analysis revealed that the production of rosmarinic acid (RA) in transformed roots of A. foeniculum was almost 4-fold higher than that of the non-transformed roots. In a separate experiment, hairy roots obtained from one-month-old leaves inoculated with A4 strain, were grown in liquid medium and the effects of different concentrations of salicylic acid (0.0, 0.01, 0.1 and 1 mM) and chitosan (0, 50, 100 and 150 mg L−1) (as elicitor) and sucrose (20, 30, 40 and 50 g L−1) on the growth of hairy roots were evaluated. The results showed that, 30 g L−1 sucrose and 100 mg L−1 chitosan increased the biomass of hairy root cultures and application of salicylic acid reduced the growth of hairy roots compared with control roots.

Technology and Application of Hairy Root Culture in Monocotyledons

2021 ◽  
Vol 07 ◽  
Author(s):  
Chang-Qi Hao ◽  
Shuai-Run Wang ◽  
Yi Wang ◽  
Xin-Yi Hou ◽  
Ya-Xuan Jiang ◽  
...  
Keyword(s):  
Medicinal Plants ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Food Crops ◽  
Root Induction ◽  
Hairy Root Induction ◽  
Selection Of

Background: Hairy root culture has been widely used in the production of metabolites in dicotyledons, and a large number of food crops and medicinal plants in monocotyledons need to be developed, but there are many difficulties in the induction of hairy roots in monocotyledons. The purpose of this paper is to introduce the inducing methods, influencing factors and application of hairy roots in monocotyledons, and to promote the development of hairy root system in monocotyledons. Methods: The mechanism of action of Agrobacterium rhizogenes and the current situation of hairy root induction, induction methods and influencing factors of monocotyledons were summarized so as to provide convenience for efficient acquisition of hairy root of monocotyledons. Results: Monocotyledons are not easy to produce phenols, cells are prone to lignification, adverse differentiation and selective response to Agrobacterium rhizogenes strains. It is proposed that before induction, plant varieties and explants should be selected, and different infection strains should be screened. In the process of hairy root induction, exogenous inducers such as acetosyringone can be added. Although these factors can provide some help for the induction of hairy roots in monocotyledons, we still need to pay attention to the disadvantages of monocotyledons from dicotyledons at the cellular level. Conclusion: A large number of food crops and medicinal plants are monocotyledons. Hairy root culture can be used to help the breeding and production of medicinal substances. Therefore, it is necessary to pay attention to the selection of varieties and explants, the selection of Agrobacterium rhizogenes and the addition of acetosyringone in the process of hairy root induction so as to improve the production efficiency and facilitate the development and utilization of monocotyledons.

scholarly journals Evaluating α-glucosidase inhibitory activity of hairy roots cultured from some plants in malvaceae family induced by Agrobacterium rhizogenes ATCC 15834

2020 ◽  
Vol 2 (6) ◽  
pp. 165-174
Author(s):  
Vu Thi Bach Phuong ◽  
Pham Thi Anh Hong ◽  
Quach Ngo Diem Phuong
Keyword(s):  
Type 2 Diabetes ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Inhibitory Activity ◽  
Root Culture ◽  
Root Induction ◽  
Hibiscus Sabdariffa ◽  
Sida Acuta

One of the most effective methods for type 2 diabetes treatments is inhibition of enzyme α-glucosidase in the intestines to slow down the release of glucose from carbohydrates in the diet, reduce plasma glucose levels and prevent hyperglycemia after meals. Therefore, seeking α-glucosidase inhibitors used in the treatment of diabetes from plant is the attention of many scientists. Based on the potential of the hairy root culture technology in increasing valuable chemical compounds accumulating, this study aimed to induce hairy roots from six plants of the Malvaceae family including Urena lobata, Abutilon indicum, Hibiscus Sabdariffa, Hibiscus rosa-sinensis, Sida acuta, Sida rhombifolia, and screening which materials has the highest in α-glucosidase inhibitory activity. We have successfully induced hairy roots from six plant species by using the Agrobacterium rhizogenes ATCC 15834 strain. The highest rates of hairy root induction were observed in Hibiscus Sabdariffa and Urena lobata. The stable introduction of rolB and rolC genes to plant genomes was confirmed by PCR. Under liquid-shake culture conditions on MS medium, hairy roots of Hibiscus sabdariffa, Urena lobata and Sida acuta showed better development than other species, and therefore, they are selected for the study of α-glucosidase inhibitory activity. This study proved that Urena lobata was stronger in inhibiting α-glucosidase activity than other studied plants, with the IC50 value of 7.65 μg/ml. The results of this study demonstrated Urena lobata hairy root might be considered as a potential supply of medicinal plants for the treatment of type 2 diabetes.  

scholarly journals Genetically transformed hairy roots producing agropine induced on Trigonella foenum-graecum L. plant by Agrobacterium rhizogenes 1601.

2013 ◽  
Vol 7 (1) ◽  
pp. 91-98
Author(s):  
M. M. Al-Mahdawe ◽  
M. K. Al-Mallah ◽  
A. O. Al-Attrakchii
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Root Formation ◽  
Paper Electrophoresis ◽  
Transformed Roots ◽  
Fresh Weight ◽  
Trigonella Foenum Graecum ◽  
Transformed Hairy Roots ◽  
Long Time ◽  
Trigonella Foenum Graecum L

Inoculation of leaves and stems explants excised from field-grown Trigonella foenum-graecum L. with engineered Agrobacterium rhizogenes 1601 inoculum of optical density 1.90, 2.06 and 1.96 led to the formation of hairy roots on these explants. The highest percent of infection with inoculum of O.D 1.90 was 20% in leaves, and was 53.3% in stems using inoculum of O.D 2.06. Infection percent was 50% in hypocotyl compared with non-inoculated explants. The results showed that inoculation of seedlings lacking roots were slow in hairy root formation and take long time compared with their formation in intact seedlings inoculated with the same inocula. Results of paper electrophoresis of these root proved the incidence of their genetic transformation. Moreover, these transformed roots grow happily in liquid WP medium with an average fresh weight 5.9g after four weeks of culture, whereas fresh weight of normal roots was 1.02g under the same conditions.

scholarly journals Hairy root culture of white mulberry (Morus alba L.) for a source of tyrosinase inhibitors

2020 ◽  
Vol 93 (1) ◽  
Author(s):  
Hanh Thi My Nguyen ◽  
Tan Quoc Tran ◽  
Anh Lan Bui ◽  
Phuong Ngo Diem Quach
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Morus Alba ◽  
Root Induction ◽  
Skin Whitening ◽  
Tyrosinase Inhibitors ◽  
White Mulberry

White mulberry (Morus alba L.) root was reported to contain potential tyrosinase inhibitors such as norartocarpetin, streppogenin, oxyresveratrol (5, 7, 2′, 4′-tetrahydroxy-flavavone) and 2-arylbenzofuran. This study aims at investigating Morus alba L. hairy root culture to create a source of tyrosinase inhibitors for skin-whitening application. Morus alba L. leaf (including mesophyll and petiole) and stem explants were randomly wounded and inoculated with Agrobacterium rhizogenes ATCC 15834 in 10-40 minutes, and coculturing for 4 days. After inoculation, Agrobacterium rhizogenes were removed by using MS medium adding 200mg/L cefotaxime as antibiotics. Then, fresh inducted hairy roots were grown in nonphytohormone liquid medium (MS, White, Gamborg B5 or WPM) on a shaker (80rpm) at 25 ±1°C. Inducted hairy roots were tested for the present of rolB by PCR analysis and its effect on tyrosinase activity using a 96-well plate reader. The suitable explants for hairy root induction are leaves, especially petioles in 10 minutes (root induction frequency: 83%). After induction, hairy roots on MS was grown more efficiently (increased 3.933 times in FW) than on WPM, B5 and White medium. Inducted hairy roots were successfully transgenes by the present of rolB in their genome and had the same anti-tyrosinase activity as the natural roots without significance differences (IC50: 3.860 and 3.597 μg/mL, respectively). Cultured Morus alba L. hairy roots are potential source of tyrosinase inhibitors for future skin-whitening ingredient.

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