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.  

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 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.

scholarly journals Hairy Root Induction on Justicia gendarussa by Various Density of Agrobacterium rhizogenes strain LB 510

2017 ◽  
Vol 9 (1) ◽  
pp. 26
Author(s):  
Dwi Kusuma Wahyuni ◽  
Siti Nur Hafida ◽  
Tri Muji Ermayanti ◽  
Bambang Prajoga Eko Wardoyo ◽  
Hery Purnobasuki ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Solid Medium ◽  
Leaf Explants ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Root Induction ◽  
Hairy Root Induction ◽  
Industrial Needs

<p>Gandarusa (<em>Justicia gendarussa</em> Burm.f.) is an Indonesian medicinal plant that has many benefits as drug and male contracetive. For industrial needs, Gandarusa must be  available in large quantity. Hairy root culture is one of methode to produce phytochemistry compound. The objective of the study was to examine the effect of various density of <em>Agrobacterium rhizogenes</em> strain LB510 on hairy roots induction of gandarusa (<em>Justicia gendarussa </em>Burm.f.) leaf plant. Leaf explants were inoculated in MS liquid medium with various density of OD<sub>600</sub> = 0.1; 0.2; 0.3; 0.4; and 0.5. Explants were co-cultivated for 2 days on MS solid medium without any hormone then sub-cultured on MS solid medium containing antibiotic cefotaxim 300 ppm, in dark condition. The data were analyzed descriptively and statistically. The results showed that various density of <em>Agrobacterium rhizogenes</em> strain LB510 was affected the lenght of hairy roots induction of <em>J</em><em>.</em><em> gendarussa</em> Burm.f., but these was not effected toward lenght formation time and number of hairy root. The treatment of OD<sub>600</sub> 0.2 was the best treatment for hairy root induction on <em>Justicia gendarussa </em>Burm. f. This data could be used for optimized the quality of methode of hairy root induction. </p>

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.

scholarly journals Comparison of Different Strains of Agrobacterium rhizogenes for Hairy Root Induction and Betulin and Betulinic Acid Production in Morus alba

2017 ◽  
Vol 12 (4) ◽  
pp. 1934578X1701200
Author(s):  
Chang Ha Park ◽  
Shicheng Zhao ◽  
Hyeon Ji Yeo ◽  
Ye Eun Park ◽  
Thanislas Bastin Baska ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Betulinic Acid ◽  
Acid Production ◽  
Leaf Tissue ◽  
Dry Weight ◽  
Morus Alba ◽  
Root Induction ◽  
Different Strains

Three Agrobacterium rhizogenes strains were tested for their ability to transform the plant Morus alba L. and to induce production of the secondary metabolites betulin and betulinic acid. All the tested strains of A. rhizogenes (R1601, LBA9402 and R1000) were able to induce hairy root formation in leaf tissue explants. Strain LBA9402 had the highest rate of infection (92.7% ± 8.8%), whereas strain R1601 had the lowest rate (87.4% ± 9.3%). The highest number of hairy roots per explant (5.6 ± 0.5) and the greatest root length (2.4 ± 0.2 mm) were obtained with strain LBA9402. We also evaluated dry weight (a measure of growth) and betulin and betulinic acid production in hairy roots and found that the highest growth (167.8 ± 14.5 mg/flask) occurred after infection with strain LBA9402. Furthermore the highest production of betulin (5.4 ± 0.4 mg/g dry weight) and betulinic acid (2.3 ± 0.2 mg/g dry weight) was noted using strain LBA9402. Among three elicitors, yeast extract showed the highest induction of betulin production (8.7 ± 0.4 mg/g) and silver nitrate induced the highest yield of betulinic acid (4.1 ± 0.2 mg/g). Our study showed that A. rhizogenes strain LBA9402 was the most effective of the three tested strains for production of transformed root cultures and betulin and betulinic acid.

scholarly journals Induction of transgenic hairy roots in soybean genotypes by Agrobacterium rhizogenes‑mediated transformation

2011 ◽  
Vol 46 (9) ◽  
pp. 1070-1075 ◽  
Author(s):  
Ricardo Luís Mayer Weber ◽  
Maria Helena Bodanese‑Zanettini
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Fluorescent Protein ◽  
Root Culture ◽  
Transformation Frequency ◽  
Plant Production ◽  
Uniform Size ◽  
Soybean Genotypes

The objective of this work was to perform the screening of soybean genotypes as to their ability to respond to the induction of hairy roots by Agrobacterium rhizogenes‑mediated transformation. Four Brazilian soybean cultivars (BRSMG 68 Vencedora, BRS 137, Embrapa 48, and MG/BR 46 Conquista) and two North American ones adapted to Brazilian cropping conditions (Bragg and IAS‑5) were screened for their capacity to respond to A. rhizogenes in protocols for in vitro hairy root culture and ex vitro composite plant production. Four‑day‑old seedlings with uniform size were injected with A. rhizogenes harboring the plasmid p35S‑GFP. Seedlings expressing green fluorescent protein (GFP) in at least one hairy root were used to determine the transformation frequency. Using an axenic in vitro protocol, excised cotyledons from four‑day‑old seedlings were infected with A. rhizogenes harboring the pCAMBIA1301 plasmid, containing the gusA reporter gene. The transformation frequency and the number of days for hairy root emergence after bacterial infection (DAI) were evaluated. The transformation frequency and DAI varied according to the genotype. Cultivars MG/BR 46 Conquista and BRSMG 68 Vencedora are more susceptible to A. rhizogenes and can be recommended for transformation experiments.

scholarly journals Effect of methyl jasmonate on the production of sesquiterpene lactones in the hairy root culture of Lactuca virosa L.

2014 ◽  
Vol 68 (2) ◽  
pp. 119-121 ◽  
Author(s):  
Janusz Malarz ◽  
Wanda Kisiel
Keyword(s):  
Methyl Jasmonate ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Hplc Analysis ◽  
Sesquiterpene Lactones ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Lactuca Virosa ◽  
Methyl Jasmonate Treatment

Hairy roots of <em>Lactuca virosa</em> L. transformed with <em>Agrobacterium rhizogenes</em> and treated with methyl jasmonate, are capable of producing sesquiterpene lactones - crepidiaside B and lactuside A in amounts higher than in untreated roots. An HPLC analysis showed the highest content of crepidiaside B after 24 hours of methyl jasmonate treatment (60% above the control). The increase in the content of lactuside A was relatively small and reached 27% after 48 hours of the treatment, in comparison with the control.

scholarly journals Agrobacterium rhizogenes Mediated Transformation of the Critically Endangered Species, Swertia chirayita

2019 ◽  
Vol 29 (2) ◽  
pp. 231-244
Author(s):  
Tapojita Samaddar ◽  
Sayantika Sarkar ◽  
Sumita Jha
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Basal Medium ◽  
Root Culture ◽  
Critically Endangered ◽  
Root Induction ◽  
Scale Production ◽  
Shoot Buds ◽  
Transformed Plants ◽  
Swertia Chirayita

Agrobacterium rhizogenes-mediated genetic transformation for the establishment of root culture lines was developed successfully in the critically endangered medicinal plant Swertia chirayita (Roxb.) H. Karst. The key factor for the successfull development of hairy root culture lines was the use of whole micropropagated plants as excised explants of any type did not show any response following infection with A. rhizogenes strains. Maximum root induction frequency (25 ± 4.4%) with 2 - 9 roots per node/intermodal cut site was obtained following infection with A. rhizogenes strain LBA9402. Spontaneous regeneration of shoot buds was observed in six 9402-transformed root lines maintained in phytohormone free N/5 basal medium. The shoot buds developed for regenerating whole rooted Ri-transformed plants in phytohormone free basal medium. Integration of rolA, rolB, rolC and rolD genes of TL-DNA was confirmed in Ri-transformed root lines and plants by PCR analysis. Ten fast growing Ri-transformed root lines were screened for production of principal secoiridoids, characteristics of the parent plants. The swertiamerin content varied from 0.042 to 0.207% in the transformed root lines after 4 weeks of culture in N/5 basal medium. Ri-transformed plants showed enhanced accumulation of swertiamerin as compared to non-transformed plants of similar age. This is the first report of swertiamerin production in hairy root cultures and transformed plants of S. chirayita thereby providing new avenue for large-scale production of secondary metabolites of S. chirayita.

scholarly journals High-Efficiency Agrobacterium rhizogenes-Mediated Transgenic Hairy Root Induction of Lens culinaris

Agronomy ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1170
Author(s):  
Chrysanthi Foti ◽  
Ourania Pavli
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Efficiency ◽  
Culture Media ◽  
Abiotic Stress Tolerance ◽  
Root Induction ◽  
Media Composition ◽  
Transgenic Hairy Root ◽  
Transgenic Hairy Roots

Previous efforts to transform lentil have been considerably hampered by the crop’s recalcitrant nature, giving rise to particularly low transformation and regeneration frequencies. This study aimed at optimizing an Agrobacterium rhizogenes-mediated transformation protocol for the generation of composite lentil plantlets, comprised of transgenic hairy roots and wild-type shoots. Transformation was performed by inoculating the cut hypocotyl of young lentil seedlings, while optimization involved the use of different bacterial strains, namely R1000, K599 and Arqua, and protocols differing in media composition with respect to the presence of acetosyringone and MES. Composite plantlets had a transgenic hairy root system characterized by an increased number of hairy roots at the hypocotyl proximal region, occasionally showing plagiotropic growth. Overall findings underline that transformation frequencies are subject to the bacterial strain, media composition as well as their combined effect. Among strains tested, R1000 proved to be the most capable of hairy root formation, while the presence of both acetosyringone and MES in inoculation and culture media yielded considerably higher transformation rates. The transgenic nature of hairy roots was demonstrated by the Ri T-DNA-mediated transfer of the rolB2 gene and the simultaneous absence of the virCD sequence of A. rhizogenes. Our findings provide strong evidence that A. rhizogenes-mediated transformation may be employed as a suitable approach for generating composite seedlings in lentil, a species whose recalcitrance severely hampers all efforts addressed to transformation and whole plant regeneration procedures. To the best of our knowledge, this is the first report on the development of a non-laborious and time-efficient protocol for the generation of transgenic hairy roots in lentil, thus providing an amenable platform for root biology and gene expression studies in the context of improving traits related to biotic and abiotic stress tolerance.

scholarly journals Induction of Platycodon grandiflorum hairy roots through the mediation of four Agrobacterium rhizogenes strains

2016 ◽  
Vol 19 (4) ◽  
pp. 64-75
Author(s):  
Phuong Dong Tra ◽  
Phuong Thi Bach Vu ◽  
Phuong Ngo Diem Quach
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Biological Activities ◽  
Root Induction ◽  
Infection Frequency ◽  
Platycodon Grandiflorum ◽  
Metabolites Production ◽  
Valuable Compounds ◽  
Agrobacterium Rhizogenes Strains

Balloon flower (Platycodon grandiflorum (Jacq.) A. DC.), the only species in Platycodon genus (Campanulaceae), is mainly distributed in East Asia. The rhizomes of P. grandiflorum, a traditional herbal medicine, have been widely used for the treatment of cough, sore throat, asthma, tuberculosis and other diseases. Recently, pharmacological researches identified important biological activities compounds in the rhizomes. Thus, to study and extract valuable compounds, a hairy root induced technique was achieved on P. grandiflorum for stable material with fast growth rates (in hormone-free media) and metabolites production. To achieve this, the “natural genetic tool” Agrobacterium rhizogenes, which can transfer DNA segments into genome of plant, was exploited. The results suggested two (A. rhizogenes ATCC 15834 and C34) of four A. rhizogenes strains could induce hairy roots. RolB and rolC genes, which are responsible for the induction of hairy roots, were inserted into the genome of hairy roots. Leaves had the highest infection frequency of hairy root induction 100 %. The optimization of protocol, including time of immersion and co-culture, had the best results with 10 and 15 mins (10 mins for A. rhizogenes ATCC 15834 and 15 mins for A. rhizogenes C34) and 72 hours, respectively. In the future, this protocol, which was described in this paper, should be useful for studying and isolating valuable compounds from P. grandiflorum hairy root cultures.

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