A Simple and Efficient Protocol for Hairy Root Culture of Arabidopsis Thaliana

2021 ◽  
Author(s):  
Pijush Paul ◽  
Sukanya Majumdar ◽  
Sumita Jha
Keyword(s):  
Arabidopsis Thaliana ◽  
Secondary Metabolites ◽  
Hairy Root ◽  
Plant Secondary Metabolites ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Bacterial Suspension ◽  
Root Induction ◽  
Role Players ◽  
High Transformation

Abstract Hairy root culture (HRC) is a valuable biotechnological tool for the production of plant secondary metabolites. Secondary metabolome study of Arabidopsis thaliana can be helpful in understanding the biological roles of various secondary metabolites present in it and HRC constructs the base of such study. The present work deals with the establishment of Agrobacterium rhizogenes strains transformed HRC of A. thaliana with very high transformation frequency resulting in long term hairy root cultures grown in hormone free media. Optimization of culture medium and standardisation of co-cultivation period are the key role players in obtaining high frequency of hairy roots. Four days of preculture in CIM medium and five min of co-cultivation in the bacterial suspension were found to be optimal conditions for root induction. This protocol could become a powerful tool for transcriptomics and proteomics-based studies for different transgenic root lines of A. thaliana.

Hairy Root Culture for In Vitro Production of Secondary Metabolites: A Promising Biotechnological Approach

2018 ◽  
pp. 235-250
Author(s):  
Ravi Shankar Singh ◽  
Tirthartha Chattopadhyay ◽  
Dharamsheela Thakur ◽  
Nitish Kumar ◽  
Tribhuwan Kumar ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Root Culture ◽  
Hairy Root Culture ◽  
In Vitro Production ◽  
Biotechnological Approach ◽  
Vitro Production

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.

Hairy Roots

2021 ◽  
pp. 219-252
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Growth ◽  
Root Culture ◽  
Hairy Root Culture ◽  
High Growth Rate ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

scholarly journals Establishment of hairy root culture and production of secondary metabolites in Coleus (Coleus forskohlii)

2014 ◽  
Vol 8 (1) ◽  
pp. 58-62 ◽  
Author(s):  
Pandey Rajiv ◽  
Krishnasamy V. ◽  
Kumaravadivel N. ◽  
Rajamani K.
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Coleus Forskohlii

scholarly journals Sugar availability modulates polyisoprenoid and phytosterol profiles in Arabidopsis thaliana hairy root culture

2013 ◽  
Vol 1831 (2) ◽  
pp. 438-447 ◽  
Author(s):  
Adam Jozwiak ◽  
Magdalena Ples ◽  
Karolina Skorupinska-Tudek ◽  
Magdalena Kania ◽  
Marta Dydak ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Hairy Root ◽  
Root Culture ◽  
Hairy Root Culture

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.

Hairy Root Culture an Alternative for Bioactive Compound Production from Medicinal Plants

2020 ◽  
Vol 22 (1) ◽  
pp. 136-149
Author(s):  
Arpita Roy
Keyword(s):  
Medicinal Plants ◽  
Secondary Metabolites ◽  
Bioactive Compounds ◽  
Hairy Root ◽  
Scale Up ◽  
Root Culture ◽  
Bioactive Compound ◽  
Hairy Root Culture ◽  
Alternative Methods ◽  
Wide Range

: Medicinal plants produce a diverse group of phytocompounds like anthraquinones, alkaloids, anthocyanins, flavonoids, saponins, and terpenes which are used in pharmaceutical, perfume, cosmetics, dye and flavor industries. Commercial source of these metabolites is field-grown plants, which are generally influenced by seasonal changes. Biotechnology possesses a significant role in production of high-value secondary metabolites. By incorporating biotechnological methods, it is feasible to manage biosynthetic pathways of the plant to enhance phytocompound production that is of pharmaceutical interest. Plant cell suspension, shoot, adventitious root and hairy root culture are considered as alternative methods for important bioactive compound production. These methods are controllable, sustainable and overcome several inconveniences for large scale secondary metabolites production. At present research on hairy root culture for valuable bioactive compound production has gained a lot of attention. Agrobacterium rhizogenes is an agent which causes hairy root disease in a plant and this leads to the neoplastic growth of root which is characterized by higher growth rate and genetic stability. Various studies explore the hairy root culture for production of a wide range of bioactive compounds. Scale-up of hairy root culture using bioreactors has provided an opportunity to enhance bioactive compound production at the commercial level. The present review discusses the role of hairy root culture in the production of valuable bioactive compounds, the effect of culture parameters on bioactive compound production and bioreactor applications.

Hairy Roots

2022 ◽  
pp. 735-759
Author(s):  
Pradip Chandra Deka
Keyword(s):  
Secondary Metabolites ◽  
Hairy Root ◽  
Hairy Roots ◽  
High Growth ◽  
Root Culture ◽  
Hairy Root Culture ◽  
High Growth Rate ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Transformed Root Cultures

Agrobacterium rhizogenes induces hairy root disease in plants. The neoplastic (cancerous) roots produced by A. rhizogenes infection, when cultured in hormone free medium, show high growth rate and genetic stability. These genetically transformed root cultures can produce levels of secondary metabolites comparable to that of intact plants. Several elicitation methods can be used to further enhance the production and accumulation of secondary metabolites. Thus, hairy root culture offer promise for high production and productivity of valuable secondary metabolites in many plants. Hairy roots can also produce recombinant proteins from transgenic roots, and thereby hold immense potential for pharmaceutical industry. Hairy root cultures can be used to elucidate the intermediates and key enzymes involved in the biosynthesis of secondary metabolites, and for phytoremediation due to their abundant neoplastic root proliferation property. Various applications of hairy root cultures and potential problems associated with them are discussed in this chapter.

scholarly journals ENHANCED ANTHOCYANIN PRODUCTION IN HAIRY ROOT CULTURE OF DAUCUS CAROTA BY FUNGAL ELICITOR

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 694b-694 ◽  
Author(s):  
Chano H. Kim ◽  
Jonq H. Park ◽  
In S. Chung ◽  
Sung R. Kim ◽  
Seung W. Lee
Keyword(s):  
Secondary Metabolites ◽  
Fluidized Bed ◽  
Hairy Root ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Fungal Elicitor ◽  
High Density Culture ◽  
Fluidized Bed Bioreactor ◽  
Fungal Elicitors ◽  
Anthocyanin Production

Secondary metabolite production by plant cell culture has been become of interest because of its commercial value in use. However, cultured plant cells usually yield lower levels of secondary metabolites than those of intact plants. In order to improve the anthocyanin productivity in hairy root culture of Daucus carota, fungal elicitors from 8 species of Fungi were examined. Through the studies of fungal elicitors in this work, it was turned out that fungal elicitors were very effective to improve the yield of anthocyanin. Despite of its low yield of anthocyanin, high density culture of hairy roots is achieved in fluidized-bed bioreactor, Anthocyanin production in fluidized-bed bioreactor with fungal elicitor treatment was increased greatly. We are currently researching more detailed aeration effects and scale-up in air-lift bioreactors. And these studies could provide important data to establish mass production system for secondary metabolites.

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